Dirofilaria and Brugia ankyrin proteins, nucleic acid molecules, and uses thereof

ABSTRACT

The present invention relates to Dirofilaria ankyrin proteins and to Brugia ankyrin proteins; to Dirofilaria ankyrin nucleic acid molecules and to Brugia ankyrin nucleic acid molecules, including those that encode such ankyrin proteins; to antibodies raised against such ankyrin proteins; and to compounds that inhibit Dirofilaria or Brugia ankyrin function. The present invention also includes methods to identify and obtain such proteins, nucleic acid molecules, antibodies, and inhibitory compounds. Also included in the present invention are therapeutic compositions comprising such proteins, nucleic acid molecules, antibodies and/or inhibitory compounds as well as the use of such therapeutic compositions to protect animals from diseases caused by parasitic helminths.

FIELD OF THE INVENTION

The present invention relates to Dirofilaria and Brugia ankyrin nucleicacid molecules, proteins encoded by such nucleic acid molecules,antibodies raised against such proteins, compounds capable of inhibitingthe function of such proteins and methods to identify such inhibitors.The present invention also includes therapeutic compositions comprisingsuch nucleic acid molecules, proteins, antibodies, and/or inhibitors, aswell as their use to protect animals from diseases caused by parasitichelminths, such as heartworm disease, elephantiasis, and hydrocele.

BACKGROUND OF THE INVENTION

Parasitic helminth infections in animals, including humans, aretypically treated by chemical drugs. One disadvantage with chemicaldrugs is that they must be administered often. For example, dogssusceptible to heartworm are typically treated monthly. Repeatedadministration of drugs, however, often leads to the development ofresistant helminth strains that no longer respond to treatment.Furthermore, many of the chemical drugs cause harmful side effects inthe animals being treated, and as larger doses become required due tothe build up of resistance, the side effects become even greater.Moreover, a number of drugs only treat symptoms of a parasitic diseasebut are unable to prevent infection by the parasitic helminth.

An alternative method to prevent parasitic helminth infection includesadministering a vaccine against a parasitic helminth. Although manyinvestigators have tried to develop vaccines based on specific antigens,it is well understood that the ability of an antigen to stimulateantibody production does not necessarily correlate with the ability ofthe antigen to stimulate an immune response capable of protecting ananimal from infection, particularly in the case of parasitic helminths.Although a number of prominent antigens have been identified in severalparasitic helminths, including in Dirofilaria and Brugia species, thereis yet to be a commercially available vaccine developed for anyparasitic helminth.

As an example of the complexity of parasitic helminths, the life cycleof D. immitis, the helminth that causes heartworm, includes a variety oflife forms, each of which presents different targets, and challenges,for immunization. In a mosquito, D. immitis microfilariae go through twolarval stages (L1 and L2) and become mature third stage larvae (L3),which can then be transmitted back to the dog when the mosquito takes ablood meal. In a dog, the L3 molt to the fourth larval stage (L4), andsubsequently to the fifth stage, or immature adults. The immature adultsmigrate to the heart and pulmonary arteries, where they mature to adultheartworms. Adult heartworms are quite large and preferentially inhabitthe heart and pulmonary arteries of an animal. Sexually mature adults,after mating, produce microfilariae which traverse capillary beds andcirculate in the vascular system of the dog. In particular, heartworm isa major problem in dogs, which typically do not develop immunity uponinfection (i.e., dogs can become reinfected even after being cured bychemotherapy). In addition, heartworm infection has been reported incats, ferrets, and humans.

As such, there remains a need to identify efficacious compositions thatprotect animals against diseases caused by parasitic helminths such asD. immitis and B. malayi. Such compositions would preferably alsoprotect animals from infection by such helminths.

The mechanisms and regulatory pathways involved in D. immitis migrationand development are not clear. From infective L3 to mature adult, thenematode has to migrate and develop, with two molts, within itsdefinitive host. It has been shown in the free living nematode,Caenorhabditis elegans (C. elegans), that the development of the larvaeis regulated by environmental signals through chemosensory neurons.Blockage of signal transmission affects the development of the nematode(Bargmann, et al., 1991, Science, 251, 1243-1246). Many neuron-relatedgenes have been identified in C. elegans. Mutations of the genes whichcontrol normal neuron function in C. elegans will not only affect thebehavior of the nematode, but will also affect the development of thelarvae and egg laying of mutated female worms. In parasitic nematodes,very little is known about mechanisms involved in the signaltransmission and the developmental regulation of the parasites. However,host and tissue specificities in parasite infections suggest thatparasitic nematodes might also need correct environmental signals fordevelopment.

Ankyrins are peripheral membrane proteins which have been found inerythrocyte, kidney and neuronal cells of mammals. Genes coding forthree different mammalian ankyrins (ankyrin_(R), ankyrin_(B) andankyrin_(G)) have been cloned. Ankyrin_(R) was originally identified aspart of the erythrocyte membrane skeleton, and was recently alsolocalized to the plasma membrane of a subpopulation of post mitoticneurons in rat brain (Lambert, et al., 1993, J. Neurosci., 13,3725-3735). Ankyrin_(B) is a developmentally regulated human brainprotein which has two alternatively spliced isoforms with molecularmasses of 220 kilodaltons (kD) and 440 kD (Kunimoto, et al., 1991, J.Cell Biology, 115, 1319-1331). Ankyrin_(G) is a more recently isolatedhuman gene that encodes two neural-specific ankyrin variants (480 kD and270 kD), which have been localized to the axonal initial segment andnode of Ranvier (Kordeli, et al., 1995, J. Biol. Chem., 270, 2352-2359).Studies on mammalian ankyrins indicate that ankyrins bind a variety ofproteins which have functions involved with the anion exchanger(Drenckhahn, et al., 1988, Science, 230, 1287-1289), Na+/K+-ATPase,amiloride-sensitive sodium channel in kidney (Smith, et al., 1991, Proc.Natl. Acad. Sci. U.S.A., 88, 6971-6975), voltage dependent sodiumchannel of the brain and the neuromuscular junction (Srinivasan, et al.,1988, Nature, 333, 177-180), and nervous system cell adhesion molecules(Davis, et al., 1994, J. Biol. Chem., 269, 27163-27166).

Analyses of mammalian ankyrins have revealed that these large proteinsare divided into three functional domains. These include an N-terminalmembrane-binding domain of about 89-95 kD, a spectrin binding domain ofabout 62 kD, and a C-terminal regulatory domain of about 50-55 kD. Themembrane-binding domain is primarily comprised of tandem repeats ofabout 33 amino acids each. This domain usually has about 22-24 copies ofthese repeats. The repeat units appear to function in binding tomembrane proteins such as anion exchangers, sodium channels, and certainadhesion molecules. The spectrin-binding domain, as the name implies,functions in binding to the spectrin-based cytoskeleton of cellspositioned inside the plasma membrane. Finally, the regulatory domain,which is the most variant domain among the different ankyrins that havebeen studied, appears to function in as a repressor and/or an activatorof the protein-binding activities of the other two domains. Some of thevariability seen in this domain among different ankyrin species appearsto be the result of alternative splicing of nascent transcripts. For areview of ankyrin structure and function, see, for example, Bennett,1992, J. Biol. Chem., 267, 8703-8706. Bennett, ibid., is incorporatedherein by reference in its entirety.

An ankyrin gene (UNC-44) has also been identified in the free livingnematode, C. elegans. Mutation of UNC-44 affects the development andfunction of the nervous system (Otsuka et al., 1995, J Cell Biology,129, 1081-1092). More recently, a cDNA encoding a 90-kilodalton (kD)neuronal protein, E1, which is reported to be an ankyrin-relatedprotein, has been cloned from the filariid nematode, Onchocera vovulus(O. volvulus), a human parasite. The cDNA was identified by usingimmuno-screening with antisera collected from putatively immuneindividuals from an endemic area of onchocerciasis. Localization studiesby immunohistochemical assay indicated that the O. volvulus E1 nativeprotein was localized to the nerve ring, the neuronal cell bodies, andthe basal labyrinth within the extracellular clefts of the hypodermis inthe adult nematode (Erttmann et al., 1996a, J. Biol. Chem., 271,1645-1650). This 462-amino acid O. volvulus protein is reported to befull length.

SUMMARY OF THE INVENTION

The present invention relates to a novel product and a process toprotect animals against parasitic helminth infection (e.g., preventand/or treat such an infection). According to the present inventionthere are provided Dirofilaria and Brugia ankyrin proteins and mimetopesthereof; Dirofilaria and Brugia ankyrin nucleic acid molecules,including those that encode such proteins; antibodies raised againstsuch ankyrin proteins (i.e., anti-Dirofilaria and anti-Brugia ankyrinantibodies); and compounds that inhibit the function of parasitichelminth ankyrins (i.e, inhibitory compounds).

The present invention also includes methods to obtain and/or identifysuch proteins, nucleic acid molecules, antibodies and inhibitorycompounds. Also included in the present invention are therapeuticcompositions comprising such proteins, nucleic acid molecules,antibodies, and/or inhibitory compounds, as well as use of suchtherapeutic compositions to protect animals from diseases caused byparasitic helminths.

One embodiment of the present invention is an isolated nucleic acidmolecule that includes either a Dirofilaria ankyrin nucleic acidmolecule, preferably a Dirofilaria immitis (D. immitis) ankyrin nucleicacid molecule, or a Brugia ankyrin nucleic acid molecule, preferably aBrugia malayi (B. malayi) ankyrin nucleic acid molecule. Such nucleicacid molecules are refcrred to as ankyrin nucleic acid molecules. A D.immitis ankyrin nucleic acid molecule preferably includes nucleic acidsequence SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ IDNO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ IDNO:13, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ IDNO:19, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ IDNO:26, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:30, SEQ IDNO:3 1, SEQ IDNO:32, SEQ ID NO:34, SEQ ID NO:35, and/or SEQ ID NO:36, and a B. malayiankyrin nucleic acid molecule preferably includes nucleic acid sequenceSEQ ID NO:37, SEQ ID NO:38, SEQ ID NO:40, and/or SEQ ID NO:41.

In one embodiment, a preferred D. immitis or B. malayi ankyrin nucleicacid molecule comprises a coding region of at least about 1500nucleotides, preferably at least about 3000 nucleotides, even morepreferably at least about 4500 nucleotides, which is capable of encodingan ankyrin protein of at least about 500 amino acids in length,preferably at least about 1000 amino acids in length, even morepreferably at least about 1500 amino acids in length. In anotherembodiment, a preferred D. imnitis or B. malayi ankyrin nucleic acidmolecule comprises a full-length coding region which encodes afull-length ankyrin protein.

The present invention also relates to recombinant molecules, recombinantviruses and recombinant cells that include an isolated ankyrin nucleicacid molecule of the present invention. Also included are methods toproduce such nucleic acid molecules, recombinant molecules, recombinantviruses and recombinant cells.

Another embodiment of the present invention includes either aDirofilaria or a Brugia ankyrin protein, or a protein that includes aDirofilaria or a Brugia ankyrin protein. Preferred ankyrin proteinsinclude D. immitis ankyrin proteins or B. malayi ankyrin proteins. Apreferred D. immitis ankyrin protein comprises amino acid sequence SEQID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:20, SEQ IDNO:23, SEQ ID NO:28, and/or SEQ ID NO:33, and a preferred B. malayiankyrin protein comprises amino acid sequence SEQ ID NO:38.

In one embodiment, a preferred D. immitis or B. malayi ankyrin proteincomprises an amino acid sequence of at least about 500 amino acids inlength, preferably at least about 1000 amino acids in length, even morepreferably at least about 1500 amino acids in length. In anotherembodiment, a preferred D. immitis or B. malayi ankyrin proteincomprises a full-length protein, i.e., a protein encoded by afull-length coding region.

The present invention also relates to: mimetopes of either Dirofilariaor Brugia ankyrin proteins, preferably to mimetopes of either D. immitisand B. malayi ankyrin proteins; isolated antibodies that selectivelybind to either Dirofilaria or Brugia ankyrin proteins or mimetopesthereof; and inhibitors of Dirofilaria or Brugia ankyrin proteinfunction. Also included are methods, including recombinant methods, toproduce proteins, mimetopes, antibodies, and inhibitors of the presentinvention.

Yet another embodiment of the present invention is a therapeuticcomposition that is capable of protecting an animal from disease causedby a parasitic helminth. Such a therapeutic composition includes one ormore of the following protective compounds: a Dirofilaria or a Brugiaankyrin protein or a mimetope thereof; an isolated Dirofilaria or Brugiaankyrin nucleic acid molecule; an isolated antibody that selectivelybinds to a Dirofilaria or a Brugia ankyrin protein; and/or a compoundcapable of inhibiting ankyrin function identified by its ability toinhibit either Dirofilaria or a Brugia ankyrin function. A preferredtherapeutic composition of the present invention also includes anexcipient, an adjuvant and/or a carrier. Preferred ankyrin nucleic acidmolecule therapeutic compositions of the present invention includegenetic vaccines, recombinant virus vaccines and recombinant cellvaccines. Also included in the present invention is a method to protectan animal from disease caused by a parasitic helminth, comprising thestep of administering to the animal a therapeutic composition of thepresent invention.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides for isolated Dirofilaria and Brugiaankyrin proteins, isolated Dirofilaria and Brugia ankyrin nucleic acidmolecules, antibodies directed against Dirofilaria and Brugia ankyrinproteins, and compounds able to inhibit parasitic helminth ankyrinfunction (i.e., inhibitory compounds). As used herein, the termsisolated Dirofilaria ankyrin proteins, isolated Brugia ankyrin proteins,isolated Dirofilaria ankyrin nucleic acid molecules, and isolated Brugiaankyrin nucleic acid molecules refers to ankyrin proteins and ankyrinnucleic acid molecules derived from parasitic helminths of the generaDirofilaria and Brugia and, as such, can be obtained from their naturalsource, or can be produced using, for example, recombinant nucleic acidtechnology or chemical synthesis. Also included in the present inventionis the use of these proteins, nucleic acid molecules, antibodies, andinhibitory compounds as therapeutic compositions to protect animals fromparasitic helminth diseases as well as in other applications, such asthose disclosed below.

The biological functions of ankyrin-related proteins in filariidnematodes are not known. However, inhibiting normal functions of thenervous system in parasitic nematodes might cause neurons to becomeinsensitive to exogenous signals and lead to defects in the developmentof the parasite. While not being bound by theory, the possible linkbetween neuronal proteins and the development of parasitic nematodesindicates that neuronal proteins, such as ankyrins, could be potentialcandidates for the development of a vaccine against parasitic nematodeinfections.

Dirofilaria and Brugia ankyrin proteins and nucleic acid molecules ofthe present invention have utility because they represent novel targetsfor anti-parasite vaccines and drugs. The products and processes of thepresent invention are advantageous because they enable the inhibition ofparasite developmental and migratory pathways that involve ankyrin.While not being bound by theory, it is believed that nematodeankyrin-like proteins might affect the development and function of thenematode nervous system, rendering neurons insensitive to exogenoussignals involved in migration and development.

Isolation of D. immitis and B. malayi ankyrin nucleic acid molecules andproteins of the present invention was surprising even in view of thereported O. volvulus E1 nucleic acid molecule and protein disclosed byErttmann, et al., 1996a, ibid., and the reported C. elegans UNC-44nucleic acid molecule and protein disclosed by Otsuka, et al., ibid. Asdescribed in more detail in the Examples, it was very difficult toisolate D. immitis ankyrin nucleic acid molecules, despite the knowledgeof these reported O. volvulus and C. elegans nucleic acid sequences.Moreover, Erttmann, et al., 1996b, Trop. Med. Int. Health, 1, 558-574,teaches away from a D. immitis analog of the O. volvulus E1 protein, inthat the reference discloses that affinity-purified rabbit antibodiesraised against the O. volvulus E1 protein do not react with D. immitisby immunohistochemical analysis (see Erttmann, et al., 1996b, ibid.,Table 2).

Furthermore, isolated D. immitis and B. malayi ankyrin nucleic acidmolecules and proteins of the present invention, and particularly D.immitis and B. malayi ankyrin nucleic acid molecules containingfull-length coding regions and full-length D. immitis and B. malayiankyrin proteins, are distinct from the O. volvulus E1 nucleic acidmolecule and protein disclosed by Erttmann, et al., 1996a, ibid. Forexample, the inventors disclose herein a D. immitis ankyrin cDNAmolecule of about 5503 nucleotides that encodes a full-length protein ofabout 1745 amino acids, and has a predicted size of about 191.7 kD. Thisnucleic acid molecule and protein are in dramatic contrast to thereported O. volvulus full-length E1 protein of 462 amino acids (i.e.,only about 26% the size of the D. immitis full-length protein), and thereported O. volvulus E1 full-length cDNA molecule of 2043 nucleotidesencoding that protein (i.e. only about 37% the size of the D. immitisfull-length nucleic acid molecule). The B. malayi ankyrin nucleic acidmolecule disclosed herein, encoding a non-full-length ankyrin protein,spans a region of the B. malayi ankyrin gene that has no similarity tothe O. volvulus E1 nucleic acid molecule and protein, i.e., the B.malayi nucleic acid molecule disclosed herein is in a region 5' to theregion that would correspond to the O. volvulus E1 nucleic acid moleculeand protein.

One embodiment of the present invention is an isolated protein thatincludes a Dirofilaria ankyrin protein or a Brugia ankyrin protein. Itis to be noted that the term "a" or "an" entity refers to one or more ofthat entity; for example, a protein refers to one or more proteins or atleast one protein. As such, the terms "a" (or "an"), "one or more" and"at least one" can be used interchangeably herein. It is also to benoted that the terms "comprising", "including", and "having" can be usedinterchangeably. According to the present invention, an isolated, orbiologically pure, protein, is a protein that has been removed from itsnatural milieu. As such, "isolated" and "biologically pure" do notnecessarily reflect the extent to which the protein has been purified.An isolated protein of the present invention can be obtained from itsnatural source, can be produced using recombinant DNA technology, or canbe produced by chemical synthesis.

As used herein, an isolated ankyrin protein of the present invention(i.e., a Dirofilaria ankyrin protein or a Brugia ankyrin protein) can bea full-length protein or any homolog of such a protein. An isolatedprotein of the present invention, including a homolog, can be identifiedin a straight-forward manner by the protein's ability to elicit animmune response against a Dirofilaria ankyrin protein or a Brugiaankyrin protein. Examples of Dirofilaria and Brugia ankyrin homologsinclude Dirofilaria and Brugia ankyrin proteins in which amino acidshave been deleted (e.g., a truncated version of the protein, such as apeptide), inserted, inverted, substituted and/or derivatized (e.g., byglycosylation, phosphorylation, acetylation, myristoylation,prenylation, palmitoylation, amidation and/or addition ofglycerophosphatidyl inositol) such that the homolog includes at leastone epitope capable of eliciting an immune response against aDirofilaria or Brugia ankyrin protein, and/or of binding to an antibodydirected against a Dirofilaria or Brugia ankyrin protein. That is, whenthe homolog is administered to an animal as an immunogen, usingtechniques known to those skilled in the art, the animal will produce animmune response against at least one epitope of a natural Dirofilaria orBrugia ankyrin protein. The ability of a protein to effect an immuneresponse can be measured using techniques known to those skilled in theart. As used herein, the term "epitope" refers to the smallest portionof a protein or other antigen capable of selectively binding to theantigen binding site of an antibody or a T-cell receptor. It is wellaccepted by those skilled in the art that the minimal size of a proteinepitope is about six to seven amino acids.

Dirofilaria and Brugia ankyrin protein homologs can be the result ofnatural allelic variation or natural mutation. Dirofilaria and Brugiaankyrin protein homologs of the present invention can also be producedusing techniques known in the art including, but not limited to, directmodifications to the protein or modifications to the gene encoding theprotein using, for example, classic or recombinant DNA techniques toeffect random or targeted mutagenesis.

Ankyrin proteins of the present invention are encoded by Dirofilariaankyrin nucleic acid molecules or Brugia ankyrin nucleic acid molecules.As used herein, a Dirofilaria or Brugia ankyrin nucleic acid moleculeincludes nucleic acid sequences related to a natural Dirofilaria orBrugia ankyrin gene, and preferably, to a D. immitis or a B. malayiankyrin gene. As used herein, a Dirofilaria or Brugia ankyrin geneincludes all regions such as regulatory regions that control productionof the Dirofilaria or Brugia ankyrin protein encoded by the gene (suchas, but not limited to, transcription, translation or post-translationcontrol regions) as well as the coding region itself, and any introns ornon-translated coding regions. As used herein, a gene that "includes" or"comprises" a sequence may include that sequence in one contiguousarray, or may include the sequence as fragmented exons. As used herein,the term "coding region" refers to a continuous linear array ofnucleotides that translates into a protein. A full-length coding regionis that coding region that is translated into a full-length, i.e., acomplete protein as would be initially translated in its naturalmillieu, prior to any post-translational modifications. In oneembodiment, a D. immitis ankyrin gene of the present invention includesthe nucleic acid sequence SEQ ID NO:32, as well as the complement of SEQID NO:32. Nucleic acid sequence SEQ ID NO:32 represents the deducedsequence of the coding strand of a cDNA (complementary DNA) denotedherein as D. immitis ankyrin nucleic acid molecule nDiAnk₅₅₀₃, theproduction of which is disclosed in the Examples. Nucleic acid moleculenDiAnk₅₅₀₃ comprises an apparently full-length coding region. Thecomplement of SEQ ID NO:32 (represented herein by SEQ ID NO:34) refersto the nucleic acid sequence of the strand complementary to the strandhaving SEQ ID NO:32, which can easily be determined by those skilled inthe art. Likewise, a nucleic acid sequence complement of any nucleicacid sequence of the present invention refers to the nucleic acidsequence of the nucleic acid strand that is complementary to (i.e., canform a double helix with) the strand for which the sequence is cited. Itshould be noted that since nucleic acid sequencing technology is notentirely error-free, SEQ ID NO:32 (as well as other nucleic acid andprotein sequences presented herein) represents an apparent nucleic acidsequence of the nucleic acid molecule encoding a D. immitis ankyrinprotein of the present invention.

In another embodiment, a D. immitis ankyrin gene or nucleic acidmolecule can be an allelic variant that includes a similar but notidentical sequence to SEQ ID NO:32, SEQ ID NO:34, or any other D.immitis nucleic acid sequence cited herein. An allelic variant of a D.immitis ankyrin gene including SEQ ID NO:32 and SEQ ID NO:34, is a genethat occurs at essentially the same locus (or loci) in the genome as thegene including SEQ ID NO:32 and SEQ ID NO:34, but which, due to naturalvariations caused by, for example, mutation or recombination, has asimilar but not identical sequence. Because natural selection typicallyselects against alterations that affect function, allelic variantsusually encode proteins having similar activity to that of the proteinencoded by the gene to which they are being compared. Allelic variantsof genes or nucleic acid molecules can also comprise alterations in the5' or 3' untranslated regions of the gene (e.g., in regulatory controlregions), or can involve alternative splicing of a nascent transcript,thereby bringing alternative exons into juxtaposition. Similarly, a B.malayi ankyrin gene or nucleic acid molecule can be an allelic variantthat includes a similar but not identical sequence to SEQ ID NO:37 andSEQ ID NO:39. Allelic variants are well known to those skilled in theart and would be expected to be found within a given parasitic helminthsuch as Dirofilaria or Brugia, since the respective genomes are diploid,and sexual reproduction will result in the reassortment of alleles.

The minimal size of an ankyrin protein homolog of the present inventionis a size sufficient to be encoded by a nucleic acid molecule capable offorming a stable hybrid (i.e., hybridize under stringent hybridizationconditions) with the complementary sequence of a nucleic acid moleculeencoding the corresponding natural protein. As used herein, "stringenthybridization conditions" refer to those experimental conditions underwhich nucleic acid molecules having similar nucleic acid sequences willanneal to each other. Stringent hybridization conditions typicallypermit the hybridization of nucleic acid molecules having at least about70% nucleic acid sequence identity with the nucleic acid molecule beingused as a probe in the hybridization reaction. Formulae to calculate theappropriate hybridization and wash conditions to achieve hybridizationpermitting 30% or less mis-match between two nucleic acid molecules aredisclosed, for example, in Meinkoth et al, 1984, Anal. Biochem 138,267-284; Meinkoth et al, ibid, is incorporated by reference herein inits entirety. As such, the size of the nucleic acid molecule encodingsuch a protein homolog is dependent on nucleic acid composition andpercent homology between the nucleic acid molecule and complementarysequence. It should also be noted that the extent of homology requiredto form a stable hybrid can vary depending on whether the homologoussequences are interspersed throughout the nucleic acid molecules or areclustered (i.e., localized) in distinct regions on the nucleic acidmolecules. The minimal size of such nucleic acid molecules is typicallyat least about 12 to about 15 nucleotides in length if the nucleic acidmolecules are GC-rich and at least about 15 to about 17 bases in lengthif they are AT-rich. As such, the minimal size of a nucleic acidmolecule used to encode an ankyrin protein homolog of the presentinvention is from about 12 to about 18 nucleotides in length. Thus, theminimal size of an ankyrin protein homolog of the present invention isfrom about 4 to about 6 amino acids in length. There is no limit, otherthan a practical limit, on the maximal size of such a nucleic acidmolecule in that the nucleic acid molecule can include a portion of agene, an entire gene, or multiple genes, or portions thereof. Thepreferred size of a protein encoded by a nucleic acid molecule of thepresent invention depends on whether a full-length, fusion, multivalent,or functional portion of such a protein is desired.

A preferred Dirofilaria or Brugia ankyrin protein of the presentinvention is a compound that when administered to an animal in aneffective manner, is capable of protecting that animal from diseasecaused by a parasitic helminth. In accordance with the presentinvention, the ability of an ankyrin protein of the present invention toprotect an animal from disease by a parasitic helminth refers to theability of that protein to, for example, treat, ameliorate and/orprevent disease caused by parasitic helminths. In one embodiment, aDirofilaria or Brugia ankyrin protein of the present invention canelicit an immune response (including a humoral and/or cellular immuneresponse) against a parasitic helminth.

Suitable parasites to target include any parasite that is essentiallyincapable of causing disease in an animal administered a Dirofilaria orBrugia ankyrin protein of the present invention. As such, parasites totarget includes any parasite that produces a protein having one or moreepitopes that can be targeted by a humoral and/or cellular immuneresponse against a Dirofilaria or Brugia ankyrin protein of the presentinvention and/or that can be targeted by an inhibitory compound thatotherwise inhibits ankyrin function (e.g., a compound that binds toankyrin thereby blocking parasite development and/or migrationregulatory pathways), thereby resulting in the decreased ability of theparasite to cause disease in an animal. Preferred parasitic helminths totarget include nematodes, cestodes, and trematodes, with nematodes beingpreferred. Preferred nematodes to target include filariid, ascarid,capillarid, strongylid, strongyloides, trichostrongyle, and trichuridnematodes. Particularly preferred nematodes are those of the generaAcanthocheilonema, Aelurostrongylus, Ancylostoma, Angiostrongylus,Ascaris, Brugia, Bunostomum, Capillaria, Chabertia, Cooperia,Crenosonia, Dictyocaulus, Dioctophyme, Dipetalonema, Diphyllobothrium,Diplydium, Dirofilaria, Dracunculus, Enterobius, Filaroides, Haemonchus,Lagochilascaris, Loa, Mansonella, Muellerius, Nanophyetus, Necator,Nematodirus, Oesophagostomum, Onchocerca, Opisthorch is, Ostertagia,Parafilaria, Paragoninius, Parascaris, Physaloptera, Protostrongylus,Setaria, Spirocerca, Spirometra, Stephanofilaria, Strongyloides,Strongylus, Thelazia, Toxascaris, Toxocara, Trichinella,Trichostrongylus, Trichuris. Unciniaria, and Wuchereria . Preferredfilariid nematodes include Dirofilaria, Onchocerca, Acanthocheilonema,Brugia, Dipetalonema, Loa, Parafilaria, Setaria, Stephanofilaria andWuchereria filariid nematodes, with D. immitis and B. malayi being evenmore preferred.

The present invention also includes mimetopes of Dirofilaria and Brugiaankyrin proteins of the present invention. As used herein, a mimetope ofa Dirofilaria or Brugia ankyrin protein of the present invention refersto any compound that is able to mimic the activity of such an ankyrinprotein, often because the mimetope has a structure that mimics theparticular ankyrin protein. Mimetopes can be, but are not limited to:peptides that have been modified to decrease their susceptibility todegradation such as all-D retro peptides; anti-idiotypic and/orcatalytic antibodies, or fragments thereof; non-proteinaceousimmunogenic portions of an isolated protein (e.g., carbohydratestructures); and synthetic or natural organic molecules, includingnucleic acids. Such mimetopes can be designed using computer-generatedstructures of proteins of the present invention. Mimetopes can also beobtained by generating random samples of molecules, such asoligonucleotides, peptides or other organic molecules, and screeningsuch samples by affinity chromatography techniques using thecorresponding binding partner.

One embodiment of a Diro filaria or Brugia ankyrin protein of thepresent invention is a fusion protein that includes a Dirofilaria orBrugia ankyrin protein-containing domain attached to one or more fusionsegments. Suitable fusion segments for use with the present inventioninclude, but are not limited to, segments that can: enhance a protein'sstability; act as an immunopotentiator to enhance an immune responseagainst a Dirofilaria or Brugia ankyrin protein; and/or assist inpurification of a Dirofilaria or Brugia ankyrin protein (e.g., byaffinity chromatography). A suitable fusion segment can be a domain ofany size that has the desired function (e.g., imparts increasedstability, imparts increased immunogenicity to a protein, and/orsimplifies purification of a protein). Fusion segments can be joined toamino and/or carboxyl termini of the Dirofilaria or Brugiaankyrin-containing domain of the protein and can be susceptible tocleavage in order to enable straight-forward recovery of a Dirofilariaor Brugia ankyrin protein. Fusion proteins are preferably produced byculturing a recombinant cell transformed with a fusion nucleic acidmolecule that encodes a protein including the fusion segment attached toeither the carboxyl and/or amino terminal end of a ankyrin-containingdomain. Preferred fusion segments include a metal binding domain (e.g.,a poly-histidine segment); an immunoglobulin binding domain (e.g.,Protein A; Protein G; T cell; B cell; Fc receptor or complement proteinantibody-binding domains); a sugar binding domain (e.g., a maltosebinding domain); and/or a "tag" domain (e.g., at least a portion ofβ-galactosidase, a strep tag peptide, a T7 tag peptide, a Flag™ peptide,or other domains that can be purified using compounds that bind to thedomain, such as monoclonal antibodies). More preferred fusion segmentsinclude metal binding domains, such as a poly-histidine segment; amaltose binding domain; a strep tag peptide, such as that available fromBiometra in Tampa, Fla.; and an S10 peptide. Examples of particularlypreferred fusion proteins of the present invention includePHIS-PDiANK₂₈₈, PHIS-PDiANK₇₈₇, PHIS-PDiANK₄₂₂, and PHIS-PDiANK₈₆₄,production of which is disclosed herein.

In another embodiment, a Dirofilaria or Brugia ankyrin protein of thepresent invention also includes at least one additional protein segmentthat is capable of protecting an animal from one or more diseases. Sucha multivalent protective protein can be produced by culturing a celltransformed with a nucleic acid molecule comprising two or more nucleicacid domains joined together in such a manner that the resulting nucleicacid molecule is expressed as a multivalent protective compoundcontaining at least two protective compounds capable of protecting ananimal from diseases caused, for example, by at least one infectiousagent.

Examples of multivalent protective compounds include, but are notlimited to, a Dirofilaria or Brugia ankyrin protein of the presentinvention attached to one or more compounds protective against one ormore other infectious agents, particularly an agent that infects humans,cats, dogs, cattle and/or horses, such as, but not limited to: viruses(e.g., adenoviruses, caliciviruses, coronaviruses, distemper viruses,hepatitis viruses, herpesviruses, immunodeficiency viruses, infectiousperitonitis viruses, leukemia viruses, oncogenic viruses, panleukopeniaviruses, papilloma viruses, parainfluenza viruses, parvoviruses, rabiesviruses, and reoviruses, as well as other cancer-causing orcancer-related viruses); bacteria (e.g., Actinomyces, Bacillus,Bacteroides, Bordetella, Bartonella, Borrelia, Brucella, Campylobacter,Capnocytophaga, Clostridium, Corynebacterium, Coxiella, Dermatophilus,Enterococcus, Ehrlichia, Escherichia, Francisella, Fusobacterium,Haemobartonella, Helicobacter, Klebsiella, L-form bacteria, Leptospira,Listeria, Mycobacteria, Mycoplasma, Neorickettsia, Nocardia,Pasteurella, Peptococcus, Peptostreptococcus, Proteus, Pseudomonas,Rickettsia, Rochalimaea, Salmonella, Shigella, Staphylococcus,Streptococcus, and Yersinia; fungi and fungal-related microorganisms(e.g., Absidia, Acremonium, Alternaria, Aspergillus, Basidiobolus,Bipolaris, Blastomyces, Candida, Chlamydia, Coccidioides, Conidiobolus,Cryptococcus, Curvalaria, Epidermophyton, Exophiala, Geotrichum,Histoplasma, Madurella, Malassezia, Microsporum, Moniliella,Mortierella, Mucor, Paecilomyces, Penicillium, Phialemonium,Phialophora, Prototheca, Pseudallescheria, Pseudomicrodochium, Pythium,Rhinosporidium, Rhizopus, Scolecobasidium, Sporothrix, Stemphylium,Trichophyton, Trichosporon, and Xylohypha; and other parasites (e.g.,Babesia, Balantidium, Besnoitia, Cryptosporidium, Eimeria,Encephalitozoon, Entamoeba, Giardia, Hammondia, Hepatozoon, Isospora,Leishmania, Microsporidia, Neospora, Nosema, Pentatrichomonas,Plasmodium, Pneumocystis, Sarcocystis, Schistosoma, Theileria,Toxoplasma, and Trypanosoma, as well as helminth parasites, such asthose disclosed herein). In one embodiment, a Dirofilaria or Brugiaankyrin protein of the present invention is attached to one or moreadditional compounds protective against heartworm disease,elephantiasis, or hydrocele. In another embodiment, one or moreprotective compounds, such as those listed above, can be included in amultivalent vaccine comprising a Dirofilaria or Brugia ankyrin proteinof the present invention and one or more other protective molecules asseparate compounds.

A preferred isolated protein of the present invention is a proteinencoded by at least one of the following nucleic acid molecules:nDiAnk₉₃₇, nDiAnk₉₃₆, nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀, nDiAnk,₁₂₂₈,nDiAnk₁₂₂₇, nDiAnk₅₇₃, nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆, nDiAnk₁₀₄₄,nDiAnk₅₅₀₃, nDiAnk₅₂₃₅, nBmAnk₉₀₈, and nBmAnk₉₀₆, or allelic variants ofany of these nucleic acid molecules. Another preferred isolated proteinis encoded by a nucleic acid molecule the having nucleic acid sequenceSEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:11, SEQ IDNO:14, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:25, SEQ IDNO:27, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:38 and/or SEQID NO:40; or an allelic variant of such a nucleic acid molecule.

Translation of SEQ ID NO:1, the coding strand of nDiAnk₉₃₇, yields aprotein of about 312 amino acids, denoted herein as PDiAnk₃₁₂, the aminoacid sequence of which is presented in SEQ ID NO:2, assuming a firstin-frame codon extending from nucleotide 1 to nucleotide 3 of SEQ IDNO:1. The coding region encoding PDiAnk₃₁₂ is presented herein asnDiAnk₉₃₆, which has the nucleotide sequence SEQ ID NO:4 (the codingstrand) and SEQ ID NO:5 (the complementary strand).

Translation of SEQ ID NO:6, the coding strand of nDiAnk₁₀₂₉, yields aprotein of about 270 amino acids, denoted herein as PDiAnk₂₇₀, the aminoacid sequence of which is presented in SEQ ID NO:7, assuming a firstin-frame codon extending from nucleotide 2 to nucleotide 4 of SEQ IDNO:6, and a termination codon extending from nucleotide 812 tonucleotide 814 of SEQ ID NO:6. The coding region encoding PDiAnk₂₇₀, notincluding the termination codon, is presented herein as nDiAnk₈₁₀, whichhas the nucleotide sequence SEQ ID NO:9 (the coding strand) and SEQ IDNO:10 (the complementary strand). The 147 amino acid residues on theC-terminal end of PDiAnk₃₁₂ lined up with 100% identity to the 147 aminoacid residues on the N-terminal end of PDiAnk₂₇₀. PDiAnk₂₇₀ thusrepresents approximately 123 amino acids of new D. immitis ankyrin aminoacid sequence. The presence of a termination codon and a poly-A tail onnDiAnk₁₀₂₉ indicates that nDiAnk₁₀₂₉ represents the authentic 3' end ofthe D. immitis ankyrin messenger RNA that was reverse-transcribed intocDNA.

Translation of SEQ ID NO:11, the coding strand of nDiAnk₆₀₀, yields aprotein of about 200 amino acids, denoted herein as PDiAnk₂₀₀, the aminoacid sequence of which is presented in SEQ ID NO:12, assuming a firstin-frame codon extending from nucleotide 1 to nucleotide 3 of SEQ IDNO:11. The 14 amino acid residues on the C-terminal end of PDiAnk₂₀₀lined up with 100% identity to the 14 amino acid residues on theN-terminal end of PDiAnk₃₁₂. PDiAnk₂₀₀ thus represents approximately 186amino acids of new D. immitis ankyrin amino acid sequence.

Translation of SEQ ID NO:14, the coding strand of nDiAnk₁₂₂₈, yields aprotein of about 409 amino acids, denoted herein as PDiAnk₄₀₉, the aminoacid sequence of which is presented in SEQ ID NO:15, assuming a firstin-frame codon extending from nucleotide 1 to nucleotide 3 of SEQ IDNO:14. The coding region encoding PDiAnk₄₀₉ is presented herein asnDiAnk₁₂₂₇, which has the nucleotide sequence SEQ ID NO:17 (the codingstrand) and SEQ ID NO:18 (the complementary strand). The 32 amino acidresidues on the C-terminal end of PDiAnk₄₀₉ lined up with 100% identityto the 32 amino acid residues on the N-terminal end of PDiAnk₂₀₀.PDiAnk₄₀₉ thus represents about 377 amino acids of new D. immitisankyrin amino acid sequence.

Translation of SEQ ID NO:19, the coding strand of nDiAnk₅₇₃, yields aprotein of about 191 amino acids, denoted herein as PDiAnk₁₉₁, the aminoacid sequence of which is presented in SEQ ID NO:20, assuming a firstin-frame codon extending from nucleotide 1 to nucleotide 3 of SEQ IDNO:19. The 32 amino acid residues on the C-terminal end of PDiAnk₁₉₁lined up with 100% identity to the 32 amino acid residues on theN-terminal end of PDiAnk₄₀₉. PDiAnk₁₉₁ thus represents approximately 159amino acids of new D. immitis ankyrin amino acid sequence.

Translation of SEQ ID NO:22, the coding strand of nDiAnk₉₁₁, yields aprotein of about 303 amino acids, denoted herein as PDiAnk₃₀₃, the aminoacid sequence of which is presented in SEQ ID NO:23, assuming a firstin-frame codon extending from nucleotide 1 to nucleotide 3 of SEQ IDNO:22. The coding region encoding PDiAnk₃₀₃ is presented herein asnDiAnk₉₀₉, which has the nucleotide sequence SEQ ID NO:25 (the codingstrand) and SEQ ID NO:26 (the complementary strand). The 54 amino acidresidues on the C-terminal end of PDiAnk₃₀₃ lined up with 100% identityto the 54 amino acid residues on the N-terminal end of the PDiAnk₄₀₉.PDiAnk₃₀₃ thus represents approximately 249 amino acids of new D.immitis ankyrin amino acid sequence.

Translation of SEQ ID NO:27, the coding strand of nDiAnk₁₀₉₆, yields aprotein of about 348 amino acids, denoted herein as PDiAnk₃₄₈, the aminoacid sequence of which is presented in SEQ ID NO:28, assuming a startcodon extending from nucleotide 51 to nucleotide 53 of SEQ ID NO:27. Thenucleic acid molecule representing the coding region encoding PDiAnk₃₄₈,denoted herein as nDiAnk₁₀₄₄, is presented herein as SEQ ID NO:30 (thecoding strand) and SEQ ID NO:31 (the complementary strand). The about 9amino acid residues on the C-terminal end of PDiAnk₃₄₈ lined up withabout 100% identity to the about 9 amino acid residues on the N-terminalend of the PDiAnk₃₀₃. PDiAnk₃₄₈ thus represents approximately 339 aminoacids of new D. immitis ankyrin amino acid sequence.

Translation of SEQ ID NO:32, the coding strand of nDiAnk₅₅₀₃, yields afull-length polypeptide of about 1745 amino acids, denoted PDiAnk₁₇₄₅,assuming a start codon extending from nucleotide 51 through nucleotide53 of SEQ ID NO:32, and a stop codon extending from nucleotide 5286through nucleotide 5285 of SEQ ID NO:32. The resulting amino acidsequence is presented as SEQ ID NO:33. The coding region encodingPDiAnk₁₇₄₅, not including the termination codon, is denoted herein asnDiANK₅₂₃₅, and has the nucleotide sequence SEQ ID NO:35 (the codingstrand) and SEQ ID NO:36 (the complementary strand). SEQ ID NO:35 ispredicted to encode a protein with a molecular mass of about 191.7 kDand with a predicted pI of about 5.76, as calculated by the DNAsisprogram (available from Hitachi Software, San Bruno, Calif.).

A homology search of a non-redundant protein database was performed withSEQ ID NO:33, using the blastp program available through the BLAST™network of the National Center for Biotechnology Information (NCBI)(National Library of Medicine, National Institutes of Health, Baltimore,Md.), available on the World Wide Web. This database includesSwissProt+PIR+SPupdate+GenPept+GPUpdate+PDB databases. The highestscoring match of the homology search at amino acid level was GenBank™accession number gil1208874, a C. elegans ankyrin-like protein, to whichSEQ ID NO:33 showed about 69% identity, spanning from about amino acid 1through about amino acid 1745 of SEQ ID NO:33. The second highesthighest scoring match of the homology search at amino acid level wasGenBank™ accession number gil406288, a human brain ankyrin protein(variant I, Ankyrin_(B)), to which SEQ ID NO:33 showed about 51%identity, spanning from about amino acid 1 through about amino acid1745.

SEQ ID NO:33 was also compared with the protein sequence of the O.volvulus E1 protein as disclosed by Erttmann, et al., 1996a, ibid. Aregion of SEQ ID NO:33 spanning from about amino acid 1282 to aboutamino acid 1745 showed about 78% identity to the 462-amino acid O.volvulus E1 protein. At the nucleotide level, the cDNA encoding SEQ IDNO:33 (i.e., SEQ ID NO:32) was compared to the cDNA encoding the O.volvulus E1 protein. A region of SEQ ID NO:32 spanning from aboutnucleotide 3423 to about nucleotide 5474 showed about 88% nucleic acididentity to the cDNA encoding the O. volvulus E1 protein.

Translation of SEQ ID NO:37, the coding strand of nBmAnk₉₀₈, yields anon-full-length polypeptide of about 302 amino acids, denoted PBmAnk₃₀₂,assuming a first inframe codon extending from nucleotide 1 throughnucleotide 3 of SEQ ID NO:37. The resulting amino acid sequence ispresented as SEQ ID NO:38. The coding region encoding PBmAnk₃₀₂ isdenoted herein as nBmANK₉₀₆, and has the nucleotide sequence SEQ IDNO:40 (the coding strand) and SEQ ID NO:41 (the complementary strand).

A homology search of a non-redundant protein database was performed onSEQ ID NO:38 using the BLAST network. The homology spans from aboutamino acid 1 through amino acid 302 of SEQ ID NO:38. The highest scoringmatch of the homology search at amino acid level was GenBank accessionnumber A57282, a C. elegans ankyrin-like protein, which was about 86%identical to SEQ ID NO:38 through a region extending from about aminoacid 353 through about amino acid 654 of A57282.

The amino acid sequence of SEQ ID NO:38 was also compared to D. immitisankyrin protein PDiAnk₁₇₄₅ (i.e. SEQ ID NO:33 of the present invention).PBmAnk₃₀₂ had 95% identity to the region of SEQ ID NO:33 spanning fromabout amino acid 341 through about amino acid 642.

Preferred ankyrin proteins of the present invention include proteinsthat are at least about 75%, preferably at least about 80%, morepreferably at least about 85%, even more preferably at least about 90%,and even more preferably at least about 95% identical to PDiAnk₁₇₄₅ ; orare at least about 90%, and preferably at least about 95%, identical toPBmAnk₃₀₂. More preferred are ankyrin proteins comprising PDiAnk₃₁₂,PDiAnk₂₇₀, PDiAnk₂₀₀, PDiAnk₄₀₉, PDiAnk₁₉₁, PDiAnk₃₀₃, PDiAnk₃₄₈,PDiAnk₁₇₄₅, or PBmAnk₃₀₂ ; and proteins enoded by alleic variants of anucleic acid molecules encoding proteins PDiAnk₃₁₂, PDiAnk₂₇₀,PDiAnk₂₀₀, PDiAnk₄₀₉, PDiAnk₁₉₁, PDiAnk₃₀₃, PDiAnk₃₄₈, PDiAnk₁₇₄₅, orPBmAnk₃₀₂.

Other preferred ankyrin proteins of the present invention includeproteins having amino acid sequences that are at least about 75%,preferably at least about 80%, more preferably at least about 85%, evenmore preferably at least about 90%, and even more preferably at leastabout 95% identical to amino acid sequence SEQ ID NO:33; or proteinshaving amino acid sequences that are at least about 90%, and preferablyat least about 95%, identical to SEQ ID NO:38. More preferred areankyrin proteins comprising amino acid sequences SEQ ID NO:2, SEQ IDNO:7, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ IDNO:28, SEQ ID NO:33 and/or SEQ ID NO:38; and ankyrin proteins encoded byallelic variants of nucleic acid molecules encoding ankyrin proteinshaving amino acid sequences SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:28, SEQ ID NO:33 and/orSEQ ID NO:38.

Particularly preferred Dirofilaria ankyrin proteins of the presentinvention comprise amino acid sequence SEQ ID NO:33 (including, but notlimited to, the proteins consisting of amino acid sequence SEQ ID NO:33,fusion proteins and multivalent proteins), and proteins encoded byallelic variants of nucleic acid molecules encoding proteins havingamino acid sequence SEQ ID NO:33; and particularly preferred Brugiaankyrin proteins of the present invention comprise amino acid sequenceSEQ ID NO:38 (including, but not limited to, the proteins consisting ofSEQ ID NO:38, fusion proteins and multivalent proteins), and proteinsencoded by allelic variants of nucleic acid molecules encoding proteinshaving amino acid sequence SEQ ID NO:38.

In one embodiment, a preferred D. immitis or B. malayi ankyrin proteinof the present invention comprises an amino acid sequence of at leastabout 500 amino acids, preferably at least about 1000 amino acids, andeven more preferably at least about 1500 amino acids. Within thisembodiment, a preferred D. inmmitis ankyrin protein of the presentinvention has an amino acid sequence comprising at least a portion ofSEQ ID NO:33. In another embodiment, a preferred D. immitis or B. malayiankyrin protein comprises a full-length protein, i.e., a protein encodedby a full-length coding region. A particularly preferred apparentlyfull-length ankyrin protein is PDiAnk₁₇₄₅.

Additional preferred ankyrin proteins of the present invention includeproteins encoded by nucleic acid molecules comprising at least a portionof nDiAnk₉₃₇, nDiAnk₉₃₆, nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀, nDiAnk₁₂₂₈,nDiAnk₁₂₂₇, nDiAnk₅₇₃, nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆, nDiAnk₁₀₄₄,nDiAnk₅₅₀₃, nDiAnk₅₂₃₅, nBmAnk₉₀₈, and/or nBmAnk₉₀₆, as well as ankyrinproteins encoded by allelic variants of such nucleic acid molecules.

Also preferred are ankyrin proteins encoded by nucleic acid moleculeshaving nucleic acid sequences comprising at least a portion of SEQ IDNO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:14,SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:22, SEQ ID NO:25, SEQ ID NO:27,SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:35, SEQ ID NO:37 and/or SEQ IDNO:40, as well as allelic variants of these nucleic acid molecules.

In another embodiment, a preferred Dirofilaria or Brugia ankyrin proteinof the present invention is encoded by a nucleic acid moleculecomprising at least about 1500 nucleotides, preferably at least about3000 nucleotides and more preferably at least about 4500 nucleotides.Within this embodiment is an ankyrin protein encoded by at least aportion nDiAnk₅₅₀₃ or by an allelic variant of this nucleic acidmolecule. In yet another embodiment, a preferred Dirofilaria or Brugiaankyrin protein of the present invention is encoded by a nucleic acidmolecule comprising an apparently full-length ankyrin coding region,i.e., a nucleic acid molecule encoding an apparently full-length ankyrinprotein.

Another embodiment of the present invention is an isolated nucleic acidmolecule comprising either a Dirofilaria ankyrin nucleic acid moleculeor a Brugia ankyrin nucleic acid molecule. The identifyingcharacteristics of such nucleic acid molecules is heretofore described.A nucleic acid molecule of the present invention can include an isolatednatural Dirofilaria or Brugia ankyrin gene or a homolog thereof, thelatter of which is described in more detail below. A nucleic acidmolecule of the present invention can include one or more regulatoryregions, full-length or partial coding regions, or combinations thereof.The minimal size of a nucleic acid molecule of the present invention isa size sufficient to allow the formation of a stable hybrid (i.e.,hybridization under stringent hybridization conditions) with thecomplementary sequence of another nucleic acid molecule. As such, theminimal size of an ankyrin nucleic acid molecule of the presentinvention is from about 12 to about 18 nucleotides in length. Preferredankyrin nucleic acid molecules include D. immitis ankyrin nucleic acidmolecules and B. malayi ankyrin nucleic acid molecules.

In accordance with the present invention, an isolated nucleic acidmolecule is a nucleic acid molecule that has been removed from itsnatural milieu (i.e., that has been subjected to human manipulation) andcan include DNA, RNA, or derivatives of either DNA or RNA. As such,"isolated" does not reflect the extent to which the nucleic acidmolecule has been purified. An isolated Dirofilaria or Brugia ankyrinnucleic acid molecule of the present invention can be isolated from itsnatural source or produced using recombinant DNA technology (e.g.,polymerase chain reaction (PCR) amplification or cloning) or chemicalsynthesis. Isolated Dirofilaria or Brugia ankyrin nucleic acid moleculescan include, for example, natural allelic variants and nucleic acidmolecules modified by nucleotide insertions, deletions, substitutions,and/or inversions in a manner such that the modifications do notsubstantially interfere with the nucleic acid molecule's ability toencode an ankyrin protein of the present invention.

A Dirofilaria or Brugia ankyrin nucleic acid molecule homolog can beproduced using a number of methods known to those skilled in the art,see, for example, Sambrook et al., 1989, Molecular Cloning: A LaboratoryManual, Cold Spring Harbor Labs Press; Sambrook et al., ibid., isincorporated by reference herein in its entirety. For example, nucleicacid molecules can be modified using a variety of techniques including,but not limited to, classic mutagenesis and recombinant DNA techniquessuch as site-directed mutagenesis, chemical treatment, restrictionenzyme cleavage, ligation of nucleic acid fragments, PCR amplification,synthesis of oligonucleotide mixtures and ligation of mixture groups to"build" a mixture of nucleic acid molecules, and combinations thereof.Nucleic acid molecule homologs can be selected by hybridization with aDirofilaria or Brugia ankyrin nucleic acid molecule or by screening thefunction of a protein encoded by the nucleic acid molecule (e.g.,ability to elicit an immune response against at least one epitope of aDirofilaria or Brugia ankyrin protein).

An isolated nucleic acid molecule of the present invention can include anucleic acid sequence that encodes at least one Dirofilaria or Brugiaankyrin protein of the present invention, examples of such proteinsbeing disclosed herein. Although the phrase "nucleic acid molecule"primarily refers to the physical nucleic acid molecule and the phrase"nucleic acid sequence" primarily refers to the sequence of nucleotideson the nucleic acid molecule, the two phrases can be usedinterchangeably, especially with respect to a nucleic acid molecule, ora nucleic acid sequence, being capable of encoding a Dirofilaria orBrugia ankyrin protein.

A preferred nucleic acid molecule of the present invention, whenadministered to an animal, is capable of protecting that animal fromdisease caused by a parasitic helminth. As will be disclosed in moredetail below, such a nucleic acid molecule can be, or encode, anantisense RNA, a molecule capable of triple helix formation, a ribozyme,or other nucleic acid-based drug compound. In additional embodiments, anucleic acid molecule of the present invention can encode a protectiveprotein (e.g., an ankyrin protein of the present invention), the nucleicacid molecule being delivered to the animal, for example, by directinjection (i.e., as a genetic vaccine) or in a vehicle such as arecombinant virus vaccine or a recombinant cell vaccine.

One embodiment of the present invention is an ankyrin nucleic acidmolecule comprising all or part of nucleic acid molecules nDiAnk₉₃₇,nDiAnk₉₃₆, nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀, nDiAnk₁₂₂₈, nDiAnk₁₂₂₇,nDiAnk₅₇₃, nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆, nDiAnk₁₀₄₄, nDiAnk₅₅₀₃,nDiAnk₅₂₃₅, nBmAnk₉₀₈, or nBmAnk₉₀₆, or allelic variants of thesenucleic acid molecules. Another preferred nucleic acid molecule of thepresent invention includes at least a portion of nucleic acid sequenceSEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:13, SEQ IDNO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ IDNO:2 1, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ IDNO:27, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ IDNO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39, SEQ IDNO:40, and/or SEQ ID NO:41, as well as allelic variants of nucleic acidmolecules having these nucleic acid sequences. Such nucleic acidmolecules can include nucleotides in addition to those included in theSEQ ID NOs, such as, but not limited to, a full-length gene, afull-length coding region, a nucleic acid molecule encoding a fusionprotein, or a nucleic acid molecule encoding a multivalent protectivecompound.

In one embodiment, an ankyrin nucleic acid molecule of the presentinvention encodes a protein that is at least about 75%, preferably atleast about 80%, more preferably at least about 85%, even morepreferably at least about 90%, and even more preferably at least about95% identical to PDiAnk₁₇₄₅ ; or is at least about 90%, and preferablyat least about 95%, identical to PBmAnk₃₀₂. Even more preferred is anucleic acid molecule encoding PDiAnk₃₁₂, PDiAnk₂₇₀, PDiAnk₂₀₀,PDiAnk₄₀₉, PDiAnk₁₉₁, PDiAnk₃₀₃, PDiAnk₃₄₈, PDiAnk₁₇₄₅, PBmAnk₃₀₂,and/or an allelic variant of such a nucleic acid molecule.

In another embodiment, an ankyrin nucleic acid molecule of the presentinvention encodes a protein having an amino acid sequence that is atleast about 75%, preferably at least about 80%, more preferably at leastabout 85%, even more preferably at least about 90%, and even morepreferably at least about 95% identical to SEQ ID NO:33; or is at leastabout 90%, and preferably at least about 95%, identical to SEQ ID NO:38.The present invention also includes an ankyrin nucleic acid moleculeencoding a protein having at least a portion of SEQ ID NO:2, SEQ IDNO:7, SEQ ID NO:12, SEQ ID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ IDNO:28, SEQ ID NO:33 and/or SEQ ID NO:38, as well as allelic variants ofan ankyrin nucleic acid molecule encoding a protein having thesesequences, including nucleic acid molecules that have been modified toaccommodate codon usage properties of the cells in which such nucleicacid molecules are to be expressed.

In another embodiment, a preferred Dirofilaria or Brugia ankyrin nucleicacid molecule encodes an ankyrin protein comprising at least about 500amino acids, preferably at least about 1000 amino acids, and morepreferably at least about 1500 amino acids; i.e., an ankyrin nucleicacid molecule that comprises a contiguous ankyrin coding region of atleast about 1500 nucleotides, preferably at least about 3000nucleotides, and more preferably at least about 4500 nucleotides.

In yet another embodiment, a preferred Dirofilaria or Brugia ankyrinnucleic acid molecule of the present invention comprises an apparentlyfull-length ankyrin coding region, i.e., the preferred nucleic acidmolecule encodes an apparently full-length ankyrin protein.

Knowing the nucleic acid sequences of certain Dirofilaria or Brugiaankyrin nucleic acid molecules of the present invention allows oneskilled in the art to, for example, (a) make copies of those nucleicacid molecules, (b) obtain nucleic acid molecules including at least aportion of such nucleic acid molecules (e.g., nucleic acid moleculesincluding full-length genes, full-length coding regions, regulatorycontrol sequences, truncated coding regions), and (c) obtain otherDirofilaria or Brugia ankyrin nucleic acid molecules. Such nucleic acidmolecules can be obtained in a variety of ways including screeningappropriate expression libraries with antibodies of the presentinvention; traditional cloning techniques using oligonucleotide probesof the present invention to screen appropriate libraries; and PCRamplification of appropriate libraries or DNA using oligonucleotideprimers of the present invention. Preferred libraries to screen or fromwhich to amplify nucleic acid molecules include Dirofilaria or BrugiaL3, L4 or adult cDNA libraries as well as genomic DNA libraries.Similarly, preferred DNA sources from which to amplify nucleic acidmolecules include Dirofilaria or Brugia L3, L4 or adult cDNA and genomicDNA. Techniques to clone and amplify genes are disclosed, for example,in Sambrook et al., ibid.

The present invention also includes nucleic acid molecules that areoligonucleotides capable of hybridizing, under stringent hybridizationconditions, with complementary regions of other, preferably longer,nucleic acid molecules of the present invention such as those comprisingDirofilaria or Brugia ankyrin nucleic acid molecules or other parasitichelminth ankyrin nucleic acid molecules. Oligonucleotides of the presentinvention can be RNA, DNA, or derivatives of either. The minimum size ofsuch oligonucleotides is the size required for formation of a stablehybrid between an oligonucleotide and a complementary sequence on anucleic acid molecule of the present invention. A preferredoligonucleotide of the present invention has a maximum size of about 100nucleotides. The present invention includes oligonucleotides that can beused as, for example, probes to identify nucleic acid molecules, primersto produce nucleic acid molecules, or therapeutic reagents to inhibitDirofilaria or Brugia ankyrin protein production or activity (e.g., asantisense-, triplex formation-, ribozyme- and/or RNA drug-basedreagents). The present invention also includes the use of sucholigonucleotides to protect animals from disease using one or more ofsuch technologies. Appropriate oligonucleotide-containing therapeuticcompositions can be administered to an animal using techniques known tothose skilled in the art.

One embodiment of the present invention includes a recombinant vector,which includes at least one isolated nucleic acid molecule of thepresent invention, inserted into any vector capable of delivering thenucleic acid molecule into a host cell. Such a vector containsheterologous nucleic acid sequences, that is nucleic acid sequences thatare not naturally found adjacent to nucleic acid molecules of thepresent invention and that preferably are derived from a species otherthan the species from which the nucleic acid molecule(s) are derived.The vector can be either RNA or DNA, either prokaryotic or eukaryotic,and typically is a virus or a plasmid. Recombinant vectors can be usedin the cloning, sequencing, and/or otherwise manipulating of Dirofilariaand Brugia ankyrin nucleic acid molecules of the present invention.

One type of recombinant vector, referred to herein as a recombinantmolecule, comprises a nucleic acid molecule of the present inventionoperatively linked to an expression vector. The phrase operativelylinked refers to insertion of a nucleic acid molecule into an expressionvector in a manner such that the molecule is able to be expressed whentransformed into a host cell. As used herein, an expression vector is aDNA or RNA vector that is capable of transforming a host cell and ofeffecting expression of a specified nucleic acid molecule. Preferably,the expression vector is also capable of replicating within the hostcell. Expression vectors can be either prokaryotic or eukaryotic, andare typically viruses or plasmids. Expression vectors of the presentinvention include any vectors that function (i.e., direct geneexpression) in recombinant cells of the present invention, including inbacterial, fungal, parasite, insect, other animal, and plant cells.Preferred expression vectors of the present invention can direct geneexpression in bacterial, yeast, helminth or other parasite, insect andmammalian cells, and more preferably in the cell types disclosed herein.

In particular, expression vectors of the present invention containregulatory sequences such as transcription control sequences,translation control sequences, origins of replication, and otherregulatory sequences that are compatible with the recombinant cell andthat control the expression of nucleic acid molecules of the presentinvention. In particular, recombinant molecules of the present inventioninclude transcription control sequences. Transcription control sequencesare sequences which control the initiation, elongation, and terminationof transcription. Particularly important transcription control sequencesare those which control transcription initiation, such as promoter,enhancer, operator and repressor sequences. Suitable transcriptioncontrol sequences include any transcription control sequence that canfunction in at least one of the recombinant cells of the presentinvention. A variety of such transcription control sequences are knownto those skilled in the art. Preferred transcription control sequencesinclude those which function in bacterial, yeast, helminth or otherendoparasite, or insect and mammalian cells, such as, but not limitedto, tac, lac, trp, trc, oxy-pro, omp/lpp, rrnB, bacteriophage lambda(such as lambda P_(L) and lambda P_(R) and fusions that include suchpromoters), bacteriophage T7, T7lac, bacteriophage T3, bacteriophageSP6, bacteriophage SP01, metallothionein, alpha-mating factor, Pichiaalcohol oxidase, alphavirus subgenomic promoter, antibiotic resistancegene, baculovirus, Heliothis zea insect virus, vaccinia virus,herpesvirus, raccoon poxvirus, other poxvirus, adenovirus,cytomegalovirus (such as immediate early promoter), simian virus 40,retrovirus, actin, retroviral long terminal repeat, Rous sarcoma virus,heat shock, phosphate and nitrate transcription control sequences aswell as other sequences capable of controlling gene expression inprokaryotic or eukaryotic cells. Additional suitable transcriptioncontrol sequences include tissue-specific promoters and enhancers aswell as lymphokine-inducible promoters (e.g., promoters inducible byinterferons or interleukins). Transcription control sequences of thepresent invention can also include naturally occurring transcriptioncontrol sequences naturally associated with parasitic helminths, such asD immitis or B. malayi transcription control sequences.

Suitable and preferred nucleic acid molecules to include in recombinantvectors of the present invention are as disclosed herein. Preferrednucleic acid molecules to include in recombinant vectors, andparticularly in recombinant molecules, include nDiAnk₉₃₇, nDiAnk₉₃₆,nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀, nDiAnk₁₂₂₈, nDiAnk₁₂₂₇, nDiAnk₅₇₃,nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆, nDiAnk₁₀₄₄, nDiAnk₅₅₀₃, nDiAnk₅₂₃₅,nBmAnk₉₀₈, and nBmAnk₉₀₆. Particularly preferred recombinant moleculesof the present invention include pTrc-nDiAnk₁₈₆₆, pTrc-nDiAnk₂₃₆₁,pTrc-nDiAnk₁₂₆₆, and pTrc-nDiAnk₈₆₄, the production of which aredescribed in the Examples section.

Recombinant molecules of the present invention may also (a) containsecretory signals (i.e., signal segment nucleic acid sequences) toenable an expressed parasitic helminth protein of the present inventionto be secreted from the cell that produces the protein and/or (b)contain fusion sequences which lead to the expression of nucleic acidmolecules of the present invention as fusion proteins. Examples ofsuitable signal segments include any signal segment capable of directingthe secretion of a protein of the present invention. Preferred signalsegments include, but are not limited to, tissue plasminogen activator(t-PA), interferon, interleukin, growth hormone, histocompatibility andviral envelope glycoprotein signal segments. Suitable fusion segmentsencoded by fusion segment nucleic acids are disclosed herein. Inaddition, a nucleic acid molecule of the present invention can be joinedto a fusion segment that directs the encoded protein to the proteosome,such as a ubiquitin fusion segment. Eukaryotic recombinant molecules mayalso include intervening and/or untranslated sequences surroundingand/or within the nucleic acid sequences of nucleic acid molecules ofthe present invention.

Another embodiment of the present invention includes a recombinant cellcomprising a host cell transformed with one or more recombinantmolecules of the present invention. Transformation of a nucleic acidmolecule into a cell can be accomplished by any method by which anucleic acid molecule can be inserted into the cell. Transformationtechniques include, but are not limited to, transfection,electroporation, microinjection, lipofection, adsorption, and protoplastfusion. A recombinant cell may remain unicellular or may grow into atissue, organ or a multicellular organism. Transformed nucleic acidmolecules of the present invention can remain extrachromosomal or canintegrate into one or more sites within a chromosome of the transformed(i.e., recombinant) cell in such a manner that their ability to beexpressed is retained. Preferred nucleic acid molecules with which totransform a cell include Dirofilaria and Brugia ankyrin nucleic acidmolecules disclosed herein. Particularly preferred nucleic acidmolecules with which to transform a cell include nDiAnk₉₃₇, nDiAnk₉₃₆,nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀, nDiAnk₁₂₂₈, nDiAnk₁₂₂₇, nDiAnk₅₇₃,nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆, nDiAnk₁₀₄₄, nDiAnk₅₅₀₃, nDiAnk₅₂₃₅,nBmAnk₉₀₈, and nBmAnk₉₀₆.

Suitable host cells to transform include any cell that can betransformed with a nucleic acid molecule of the present invention. Hostcells can be either untransformed cells or cells that are alreadytransformed with at least one nucleic acid molecule (e.g., nucleic acidmolecules encoding one or more proteins of the present invention and/orother proteins useful in the production of multivalent vaccines). Hostcells of the present invention either can be endogenously (i.e.,naturally) capable of producing Dirofilaria or Brugia ankyrin proteinsof the present invention or can be capable of producing such proteinsafter being transformed with at least one nucleic acid molecule of thepresent invention. Host cells of the present invention can be any cellcapable of producing at least one protein of the present invention, andinclude bacterial, fungal (including yeast), parasite (includinghelminth, protozoa and ectoparasite), other insect, other animal andplant cells. Preferred host cells include bacterial, mycobacterial,yeast, helminth, insect and mammalian cells. More preferred host cellsinclude Salmonella, Escherichia, Bacillus, Listeria, Saccharomyces,Spodoptera, Mycobacteria, Trichoplusia, BHK (baby hamster kidney) cells,MDCK cells (Madin-Darby canine kidney cell line), CRFK cells (Crandellfeline kidney cell line), CV-1 cells (African monkey kidney cell lineused, for example, to culture raccoon poxvirus), COS (e.g., COS-7)cells, and Vero cells. Particularly preferred host cells are Escherichiacoli, including E. coli K-12 derivatives; Salmonella typhi; Salmonellatyphimurium, including attenuated strains such as UK-1 _(X) 3987 andSR-11 _(X) 4072; Spodoptera frugiperda; Trichoplusia ni; BHK cells; MDCKcells; CRFK cells; CV-1 cells; COS cells; Vero cells; andnon-tumorigenic mouse myoblast G8 cells (e.g., ATCC CRL 1246).Additional appropriate mammalian cell hosts include other kidney celllines, other fibroblast cell lines (e.g., human, murine or chickenembryo fibroblast cell lines), myeloma cell lines, Chinese hamster ovarycells, mouse NIH/3T3 cells, LMTK³¹ cells and/or HeLa cells. In oneembodiment, the proteins may be expressed as heterologous proteins inmyeloma cell lines employing immunoglobulin promoters.

A recombinant cell is preferably produced by transforming a host cellwith one or more recombinant molecules, each comprising one or morenucleic acid molecules of the present invention operatively linked to anexpression vector containing one or more transcription controlsequences, examples of which are disclosed herein. Particularlypreferred recombinant molecules include pTrc-nDiAnk₁₈₆₆,pTrc-nDiAnk₂₃₆₁, pTrc-nDiAnk₁₂₆₆, and pTrc-nDiAnk₈₆₄.

A recombinant cell of the present invention includes any celltransformed with at least one of any nucleic acid molecule of thepresent invention. Suitable and preferred nucleic acid molecules as wellas suitable and preferred recombinant molecules with which to transfercells are disclosed herein. Particularly preferred recombinant cellsinclude E. coli:pTrc-nDiAnk₁₈₆₆, E. coli:pTrc-nDiAnk₂₃₆₁, E.coli:pTrc-nDiAnk₁₂₆₆, and E. coli:pTrc-nDiAnk₈₆₄. Details regarding theproduction of these recombinant cells are disclosed herein.

Recombinant cells of the present invention can also be co-transformedwith one or more recombinant molecules including Dirofilaria or Brugiaankyrin nucleic acid molecules encoding one or more proteins of thepresent invention and one or more other nucleic acid molecules encodingother protective compounds, as disclosed herein (e.g., to producemultivalent vaccines).

Recombinant DNA technologies can be used to improve expression oftransformed nucleic acid molecules by manipulating, for example, thenumber of copies of the nucleic acid molecules within a host cell, theefficiency with which those nucleic acid molecules are transcribed, theefficiency with which the resultant transcripts are translated, and theefficiency of post-translational modifications. Recombinant techniquesuseful for increasing the expression of nucleic acid molecules of thepresent invention include, but are not limited to, operatively linkingnucleic acid molecules to high-copy number plasmids, integration of thenucleic acid molecules into one or more host cell chromosomes, additionof vector stability sequences to plasmids, substitutions ormodifications of transcription control signals (e.g., promoters,operators, enhancers), substitutions or modifications of translationalcontrol signals (e.g., ribosome binding sites, Shine-Dalgarnosequences), modification of nucleic acid molecules of the presentinvention to correspond to the codon usage of the host cell, deletion ofsequences that destabilize transcripts, and use of control signals thattemporally separate recombinant cell growth from recombinant enzymeproduction during fermentation. The activity of an expressed recombinantprotein of the present invention may be improved by fragmenting,modifying, or derivatizing nucleic acid molecules encoding such aprotein.

Isolated Dirofilaria or Brugia ankyrin proteins of the present inventioncan be produced in a variety of ways, including production and recoveryof natural proteins, production and recovery of recombinant proteins,and chemical synthesis of the proteins. In one embodiment, an isolatedprotein of the present invention is produced by culturing a cell capableof expressing the protein under conditions effective to produce theprotein, and recovering the protein. A preferred cell to culture is arecombinant cell of the present invention. Effective culture conditionsinclude, but are not limited to, effective media, bioreactor,temperature, pH and oxygen conditions that permit protein production. Aneffective, medium refers to any medium in which a cell is cultured toproduce a Dirofilaria or Brugia ankyrin protein of the presentinvention. Such medium typically comprises an aqueous medium havingassimilable carbon, nitrogen and phosphate sources, and appropriatesalts, minerals, metals and other nutrients, such as vitamins. Cells ofthe present invention can be cultured in conventional fermentationbioreactors, shake flasks, test tubes, microtiter dishes, and petriplates. Culturing can be carried out at a temperature, pH and oxygencontent appropriate for a recombinant cell. Such culturing conditionsare within the expertise of one of ordinary skill in the art. Examplesof suitable conditions are included in the Examples section.

Depending on the vector and host system used for production, resultantproteins of the present invention may either remain within therecombinant cell; be secreted into the fermentation medium; be secretedinto a space between two cellular membranes, such as the periplasmicspace in E. coli; or be retained on the outer surface of a cell or viralmembrane.

The phrase "recovering the protein", as well as similar phrases, refersto collecting the whole fermentation medium containing the protein andneed not imply additional steps of separation or purification. Proteinsof the present invention can be purified using a variety of standardprotein purification techniques, such as, but not limited to, affinitychromatography, ion exchange chromatography, filtration,electrophoresis, hydrophobic interaction chromatography, gel filtrationchromatography, reverse phase chromatography, concanavalin Achromatography, chromatofocusing and differential solubilization.Proteins of the present invention are preferably retrieved in"substantially pure" form. As used herein, "substantially pure" refersto a purity that allows for the effective use of the protein as atherapeutic composition or diagnostic. A therapeutic composition foranimals, for example, should exhibit no substantial toxicity andpreferably should be capable of stimulating the production of antibodiesin a treated animal.

The present invention also includes isolated (i.e., removed from theirnatural milieu) antibodies that selectively bind to a Dirofilaria orBrugia ankyrin protein of the present invention or a mimetope thereof(e.g., anti-Dirofilaria ankyrin antibodies or anti-Brugia ankyrinantibodies). As used herein, the term "selectively binds to" an ankyrinprotein refers to the ability of antibodies of the present invention topreferentially bind to specified proteins and mimetopes thereof of thepresent invention. Binding can be measured using a variety of methodsstandard in the art including enzyme immunoassays (e.g., ELISA),immunoblot assays, etc.; see, for example, Sambrook et al., ibid., andHarlow, et al., 1988, Antibodies, a Laboratory Manual, Cold SpringHarbor Labs Press; Harlow et al., ibid., is incorporated by referenceherein in its entirety. An anti-ankyrin antibody of the presentinvention preferably selectively binds to a Dirofilaria or Brugiaankyrin protein in such a way as to inhibit the function of thatprotein.

Isolated antibodies of the present invention can include antibodies inserum, or antibodies that have been purified to varying degrees.Antibodies of the present invention can be polyclonal or monoclonal, orcan be functional equivalents such as antibody fragments andgenetically-engineered antibodies, including single chain antibodies orchimeric antibodies that can bind to one or more epitopes.

A preferred method to produce antibodies of the present inventionincludes (a) administering to an animal an effective amount of aprotein, peptide or mimetope thereof of the present invention to producethe antibodies and (b) recovering the antibodies. In another method,antibodies of the present invention are produced recombinantly usingtechniques as heretofore disclosed to produce ankyrin proteins of thepresent invention. Antibodies raised against defined proteins ormimetopes can be advantageous because such antibodies are notsubstantially contaminated with antibodies against other substances thatmight otherwise cause interference in a diagnostic assay or side effectsif used in a therapeutic composition.

Antibodies of the present invention have a variety of potential usesthat are within the scope of the present invention. For example, suchantibodies can be used (a) as therapeutic compounds to passivelyimmunize an animal in order to protect the animal from parasitichelminths susceptible to treatment by such antibodies, (b) as reagentsin assays to detect infection by such helminths and/or (c) as tools toscreen expression libraries and/or to recover desired proteins of thepresent invention from a mixture of proteins and other contaminants.Furthermore, antibodies of the present invention can be used to targetcytotoxic agents to parasitic helminths in order to directly kill suchhelminths. Targeting can be accomplished by conjugating (i.e., stablyjoining) such antibodies to the cytotoxic agents using techniques knownto those skilled in the art. Suitable cytotoxic agents are known tothose skilled in the art.

One embodiment of the present invention is a therapeutic compositionthat, when administered to an animal in an effective manner, is capableof protecting that animal from disease caused by a parasitic helminth.Therapeutic compositions of the present invention include at least oneof the following protective compounds: an isolated Dirofilaria or Brugiaankyrin protein or a mimetope thereof, an isolated Dirofilaria or Brugiaankyrin nucleic acid molecule, an isolated antibody that selectivelybinds to a Dirofilaria or Brugia ankyrin protein, an inhibitor ofankyrin function identified by its ability to bind to a Dirofilaria orBrugia ankyrin protein and thereby impede development and/or migrationof the parasite, and a mixture thereof (i.e., combination of at leasttwo of the compounds). As used herein, a protective compound refers to acompound that, when administered to an animal in an effective manner, isable to treat, ameliorate, and/or prevent disease caused by a parasitichelminth. Preferred helminths to target are heretofore disclosed.Examples of proteins, nucleic acid molecules, antibodies and inhibitorsof the present invention are disclosed herein.

The present invention also includes a therapeutic composition comprisingat least one Dirofilaria or Brugia ankyrin-based compound of the presentinvention in combination with at least one additional compoundprotective against one or more infectious agents. Examples of suchcompounds and infectious agents are disclosed herein.

Therapeutic compositions of the present invention can be administered toany animal susceptible to such therapy, preferably to mammals, and morepreferably to dogs, cats, humans, ferrets, horses, cattle, sheep andother pets, economic food animals and/or zoo animals. Preferred animalsto protect against heartworm disease include dogs, cats, humans andferrets, with dogs and cats being particularly preferred. The preferredanimals to protect against elephantiasis and hydrocele include humans.

In one embodiment, a therapeutic composition of the present inventioncan be administered to the vector in which the parasitic helminthdevelops, such as to a mosquito in order to prevent the spread ofheartworm. Such administration could be oral or by developing transgenicvectors capable of producing at least one therapeutic composition of thepresent invention. In another embodiment, an insect vector, such as amosquito, can ingest therapeutic compositions present in the blood of ahost that has been administered a therapeutic composition of the presentinvention.

In order to protect an animal from disease caused by a parasitichelminth, a therapeutic composition of the present invention isadministered to the animal in an effective manner such that thecomposition is capable of protecting that animal from a disease causedby a parasitic helminth. Therapeutic compositions of the presentinvention can be administered to animals prior to infection in order toprevent infection (i.e., as a preventative vaccine) and/or can beadministered to animals after infection in order to treat disease causedby the parasitic helminth (i.e., as a therapeutic vaccine).

Therapeutic compositions of the present invention can be formulated inan excipient that the animal to be treated can tolerate. Examples ofsuch excipients include water, saline, Ringer's solution, dextrosesolution, Hank's solution, and other aqueous physiologically balancedsalt solutions. Nonaqueous vehicles, such as fixed oils, sesame oil,ethyl oleate, or triglycerides may also be used. Other usefulformulations include suspensions containing viscosity enhancing agents,such as sodium carboxymethylcellulose, sorbitol, or dextran. Excipientscan also contain minor amounts of additives, such as substances thatenhance isotonicity and chemical stability. Examples of buffers includephosphate buffer, bicarbonate buffer and Tris buffer, while examples ofpreservatives include thimerosal, m- or o-cresol, formalin and benzylalcohol. Standard formulations can either be liquid injectables orsolids which can be taken up in a suitable liquid as a suspension orsolution for injection. Thus, in a non-liquid formulation, the excipientcan comprise dextrose, human serum albumin, preservatives, etc., towhich sterile water or saline can be added prior to administration.

In one embodiment of the present invention, a therapeutic compositioncan include an adjuvant. Adjuvants are agents that are capable ofenhancing the immune response of an animal to a specific antigen.Suitable adjuvants include, but are not limited to, cytokines,chemokines, and compounds that induce the production of cytokines andchemokines (e.g., granulocyte macrophage colony stimulating factor(GM-CSF), granulocyte colony stimulating factor (G-CSF), macrophagecolony stimulating factor (M-CSF), colony stimulating factor (CSF),erythropoietin (EPO), interleukin 2 (IL-2), interleukin-3 (IL-3),interleukin 4 (IL-4), interleukin 5 (IL-5), interleukin 6 (IL-6)interleukin 7 (IL-7), interleukin 8 (IL-8), interleukin 10 (IL-10),interleukin 12 (IL-12), interferon gamma, interferon gamma inducingfactor I (IGIF), transforming growth factor beta, RANTES (regulated uponactivation, normal T-cell expressed and presumably secreted), macrophageinflammatory proteins (e.g., MIP-1 alpha and MIP-1 beta), and Leishmaniaelongation initiating factor (LEIF)); bacterial components (e.g.,endotoxins, in particular superantigens, exotoxins and cell wallcomponents); aluminum-based salts; calcium-based salts; silica;polynucleotides; toxoids; serum proteins, viral coat proteins; blockcopolymer adjuvants (e.g., Hunter's Titermax™ adjuvant (Vaxcel™, Inc.Norcross, GA), Ribi adjuvants (Ribi ImmunoChem Research, Inc., Hamilton,Mont.); and saponins and their derivatives (e.g., Quil A (SuperfosBiosector A/S, Denmark). Protein adjuvants of the present invention canbe delivered in the form of the protein themselves or of nucleic acidmolecules encoding such proteins using the methods described herein.

In one embodiment of the present invention, a therapeutic compositioncan include a carrier. Carriers include compounds that increase thehalf-life of a therapeutic composition in the treated animal. Suitablecarriers include, but are not limited to, polymeric controlled releasevehicles, biodegradable implants, liposomes, bacteria, viruses, othercells, oils, esters, and glycols.

One embodiment of the present invention is a controlled releaseformulation that is capable of slowly releasing a composition of thepresent invention into an animal. As used herein, a controlled releaseformulation comprises a composition of the present invention in acontrolled release vehicle. Suitable controlled release vehiclesinclude, but are not limited to, biocompatible polymers, other polymericmatrices, capsules, microcapsules, microparticles, bolus preparations,osmotic pumps, diffusion devices, liposomes, lipospheres, andtransdermal delivery systems. Other controlled release formulations ofthe present invention include liquids that, upon administration to ananimal, form a solid or a gel in situ. Preferred controlled releaseformulations are biodegradable (i.e., bioerodible).

A preferred controlled release formulation of the present invention iscapable of releasing a composition of the present invention into theblood of the treated animal at a constant rate sufficient to attaintherapeutic dose levels of the composition to protect an animal fromdisease caused by parasitic helminths. The therapeutic composition ispreferably released over a period of time ranging from about 1 to about12 months. A controlled release formulation of the present invention iscapable of effecting a treatment preferably for at least about 1 month,more preferably for at least about 3 months, even more preferably for atleast about 6 months, even more preferably for at least about 9 months,and even more preferably for at least about 12 months.

Therapeutic compositions of the present invention can be administered toanimals prior to infection in order to prevent infection and/or can beadministered to animals after infection in order to treat disease causedby the parasitic helminth. For example, proteins, mimetopes thereof, andantibodies thereof can be used as immunotherapeutic agents. Acceptableprotocols to administer therapeutic compositions in an effective mannerinclude individual dose size, number of doses, frequency of doseadministration, and mode of administration. Determination of suchprotocols can be accomplished by those skilled in the art. A suitablesingle dose is a dose that is capable of protecting an animal fromdisease when administered one or more times over a suitable time period.For example, a preferred single dose of a protein, mimetope or antibodytherapeutic composition is from about 1 microgram (μg) to about 10milligrams (mg) of the therapeutic composition per kilogram body weightof the animal. Booster vaccinations can be administered from about 2weeks to several years after the original administration. Boosteradministrations preferably are administered when the immune response ofthe animal becomes insufficient to protect the animal from disease. Apreferred administration schedule is one in which from about 10 μg toabout 1 mg of the therapeutic composition per kg body weight of theanimal is administered from about one to about two times over a timeperiod of from about 2 weeks to about 12 months. Modes of administrationcan include, but are not limited to, subcutaneous, intradermal,intravenous, intranasal, oral, transdermal and intramuscular routes.

According to one embodiment, a nucleic acid molecule of the presentinvention can be administered to an animal in a fashion to enableexpression of that nucleic acid molecule into a protective protein orprotective RNA (e.g., antisense RNA, ribozyme, triple helix forms or RNAdrug) in the animal. Nucleic acid molecules can be delivered to ananimal in a variety of methods including, but not limited to, (a)administering a naked (i.e., not packaged in a viral coat or cellularmembrane) nucleic acid as a genetic vaccine (e.g., as naked DNA or RNAmolecules, such as is taught, for example in Wolff et al., 1990, Science247, 1465-1468) or (b) administering a nucleic acid molecule packaged asa recombinant virus vaccine or as a recombinant cell vaccine (i.e., thenucleic acid molecule is delivered by a viral or cellular vehicle).

A genetic (i.e., naked nucleic acid) vaccine of the present inventionincludes a nucleic acid molecule of the present invention and preferablyincludes a recombinant molecule of the present invention that preferablyis replication, or otherwise amplification, competent. A genetic vaccineof the present invention can comprise one or more nucleic acid moleculesof the present invention in the form of, for example, a dicistronicrecombinant molecule. Preferred genetic vaccines include at least aportion of a viral genome (i.e., a viral vector). Preferred viralvectors include those based on alphaviruses, poxviruses, adenoviruses,herpesviruses, picornaviruses, and retroviruses, with those based onalphaviruses (such as sindbis or Semliki forest virus), species-specificherpesviruses and poxviruses being particularly preferred. Any suitabletranscription control sequence can be used, including those disclosed assuitable for protein production. Particularly preferred transcriptioncontrol sequences include cytomegalovirus immediate early (preferably inconjunction with Intron-A), Rous sarcoma virus long terminal repeat, andtissue-specific transcription control sequences, as well astranscription control sequences endogenous to viral vectors if viralvectors are used. The incorporation of a "strong" polyadenylation signalis also preferred.

Genetic vaccines of the present invention can be administered in avariety of ways, with intramuscular, subcutaneous, intradermal,transdermal, intranasal and oral routes of administration beingpreferred. A preferred single dose of a genetic vaccine ranges fromabout 1 nanogram (ng) to about 600 μg, depending on the route ofadministration and/or method of delivery, as can be determined by thoseskilled in the art. Suitable delivery methods include, for example, byinjection, as drops, aerosolized and/or topically. Genetic vaccines ofthe present invention can be contained in an aqueous excipient (e.g.,phosphate buffered saline) alone or in a carrier (e.g., lipid-basedvehicles).

A recombinant virus vaccine of the present invention includes arecombinant molecule of the present invention that is packaged in aviral coat and that can be expressed in an animal after administration.Preferably, the recombinant molecule is packaging- orreplication-deficient and/or encodes an attenuated virus. A number ofrecombinant viruses can be used, including, but not limited to, thosebased on alphaviruses, poxviruses, adenoviruses, herpesviruses,picornaviruses, and retroviruses. Preferred recombinant virus vaccinesare those based on alphaviruses (such as Sindbis virus), raccoonpoxviruses, species-specific herpesviruses and species-specificpoxviruses. An example of methods to produce and use alphavirusrecombinant virus vaccines are disclosed in PCT Publication No. WO94/17813, by Xiong et al., published Aug. 18, 1994, which isincorporated by reference herein in its entirety.

When administered to an animal, a recombinant virus vaccine of thepresent invention infects cells within the immunized animal and directsthe production of a protective protein or RNA nucleic acid molecule thatis capable of protecting the animal from disease caused by a parasitichelminth as disclosed herein. For example, a recombinant virus vaccinecomprising an ankyrin nucleic acid molecule of the present invention isadministered according to a protocol that results in the animalproducing a sufficient immune response to protect itself from heartworm.A preferred single dose of a recombinant virus vaccine of the presentinvention is from about 1×10⁴ to about 1×10⁸ virus plaque forming units(pfu) per kilogram body weight of the animal. Administration protocolsare similar to those described herein for protein-based vaccines, withsubcutaneous, intramuscular, intranasal and oral administration routesbeing preferred.

A recombinant cell vaccine of the present invention includes recombinantcells of the present invention that express at least one protein of thepresent invention. Preferred recombinant cells for this embodimentinclude Salmonella, E. coli, Listeria, Mycobacterium, S. frugiperda,yeast, (including Saccharomyces cerevisiae and Pichia pastoris), BHK,CV-1, myoblast G8, COS (e.g., COS-7), Vero, MDCK and CRFK recombinantcells. Recombinant cell vaccines of the present invention can beadministered in a variety of ways but have the advantage that they canbe administered orally, preferably at doses ranging from about 10⁸ toabout 10¹² cells per kilogram body weight. Administration protocols aresimilar to those described herein for protein-based vaccines.Recombinant cell vaccines can comprise whole cells, cells stripped ofcell walls or cell lysates.

The efficacy of a therapeutic composition of the present invention toprotect an animal from disease caused by a parasitic helminth can betested in a variety of ways including, but not limited to, detection ofprotective antibodies (using, for example, proteins or mimetopes of thepresent invention), detection of cellular immunity within the treatedanimal, or challenge of the treated animal with the parasitic helminthto determine whether the treated animal is resistant to disease.Challenge studies can include implantation of chambers includingparasitic helminth larvae into the treated animal and/or directadministration of larvae to the treated animal. In one embodiment,therapeutic compositions can be tested in animal models such as mice.Such techniques are known to those skilled in the art.

One preferred embodiment of the present invention is the use ofDirofilaria or Brugia ankyrin proteins, nucleic acid molecules,antibodies and inhibitors of the present invention, to protect an animalfrom heartworm. It is particularly preferred to prevent L3 that aredelivered to the animal by the mosquito intermediate host from migratingfrom the site of inoculation and/or maturing into adult worms. As such,preferred therapeutic compositions are those that are able to inhibit atleast one step in the portion of the parasite's development cycle thatincludes L3, third molt, L4, fourth molt, immature adult prior toentering the circulatory system. In dogs, this portion of thedevelopment cycle is about 70 days. Particularly preferred therapeuticcompositions include D. immitis ankyrin-based therapeutic compositionsof the present invention. Such compositions include D. immitis ankyrinnucleic acid molecules, D. immitis ankyrin proteins and mimetopesthereof, anti-D. immitis ankyrin antibodies, and inhibitors of D.immitis ankyrin function. Therapeutic compositions are administered toanimals in a manner effective to protect the animals from heartworm.Additional protection may be obtained by administering additionalprotective compounds, including other parasitic helminth proteins,nucleic acid molecules, antibodies and inhibitory compounds, asdisclosed herein.

One therapeutic composition of the present invention includes aninhibitor of Dirofilaria or Brugia ankyrin function, i.e., a compoundcapable of substantially interfering with the function of a Dirofilariaor Brugia ankyrin protein susceptible to inhibition. For example, anisolated protein or mimetope thereof is administered in an amount andmanner that elicits (i.e., stimulates) an immune response that issufficient, upon interaction with a native ankyrin protein, to protectthe animal from the disease. Similarly, an antibody of the presentinvention, when administered to an animal in an effective manner, isadministered in an amount so as to be present in the animal at a titerthat is sufficient, upon interaction of that antibody with a nativeankyrin protein, to protect the animal from the disease, at leasttemporarily. Oligonucleotide nucleic acid molecules of the presentinvention can also be administered in an effective manner, therebyreducing expression of Dirofilaria or Brugia ankyrin proteins in orderto interfere with development of parasitic helminths targeted inaccordance with the present invention. Methods to identify certainfunctions, i.e., protein-binding capabilities, of Dirofilaria or Brugiaankyrin proteins of the present invention are disclosed herein.

One embodiment of the present invention is a method to identify proteinsthat specifically interact with an ankyrin protein of the presentinvention. The method can comprise the steps of a) identifying andisolating a protein-binding domain of an isolated Dirofilaria or Brugiaankyrin protein; b) contacting that protein-binding domain with isolatedparasitic helminth proteins under conditions such that a parasitichelminth protein and the protein-binding domain can selectively interactand/or bind to each other, using, for example, the yeast two-hybridsystem see, for example, Luban, et al., 1995, Curr. Opin. Biotechnol.,6, 59-64; and c) identifying those proteins that specifically bind tothe isolated ankyrin protein-binding domain. Additional methods toidentify protein-protein interactions with the protein-binding domainsof an isolated ankyrin protein of the present invention are known tothose skilled in the art. Examples include Biacore® screening, confocalimmunofluorescent microscopy, and immunoprecipitations.

An inhibitor of ankyrin function can be identified using Dirofilaria orBrugia ankyrin proteins of the present invention. One embodiment of thepresent invention is a method to identify a compound capable ofinhibiting ankyrin function of a parasitic helminth. Such a methodincludes the steps of: (a) identifying a protein binding or regulatoryactivity of an isolated ankyrin protein in vitro; (b) identifying aputative compound capable of binding to and/or inhibiting the identifiedprotein binding or regulatory activity of the isolated ankyrin protein;(c) contacting D. immitis L3 larvae with the putative inhibitorycompound under conditions in which, in the absence of the compound, thelarvae are able to molt to the L4 stage; and (d) determining if theputative compound inhibits molting. Putative inhibitory compounds toscreen include small organic molecules, antibodies (including mimetopesthereof), and ligand analogs. Such compounds are also screened toidentify those that are substantially not toxic in host animals.

Inhibitors of ankyrin function identified by such a method can be testedfor their ability to block development and/or migration of parasitichelminths, and particularly of D. immitis and B. malayi, in vivo.Preferred ankyrin proteins to inhibit are those produced by parasitichelminths, even more preferred ankyrin proteins to inhibit are thoseproduced by filariid nematodes. A particularly preferred inhibitor ofthe present invention is capable of protecting an animal from heartwormdisease, elephantiasis and/or hydrocele. It is also within the scope ofthe present invention to use inhibitors of the present invention totarget diseases caused by parasitic helminths in animals. Compositionscomprising inhibitors of ankyrin function can be administered to animalsin an effective manner to protect animals from disease caused byparasitic helminths, and preferably to protect animals from heartwormdisease, elephantiasis and/or hydrocele. Effective amounts and dosingregimens can be determined using techniques known to those skilled inthe art.

It is also within the scope of the present invention to use isolatedproteins, mimetopes, nucleic acid molecules and antibodies of thepresent invention as diagnostic reagents to detect infection byparasitic helminths. Such diagnostic reagents can be supplemented withadditional compounds that can specifically detect all phases of theparasite's life cycle. Methods to use such diagnostic reagents todiagnose parasitic helminth infection are well known to those skilled inthe art. Suitable and preferred parasitic helminths to detect are thoseto which therapeutic compositions of the present invention are targeted.Particularly preferred parasitic helminths to detect using diagnosticreagents of the present invention are D. immitis and B. malayi.

The following examples are provided for the purposes of illustration andare not intended to limit the scope of the present invention.

EXAMPLES

It is to be noted that the examples include a number of molecularbiology, microbiology, immunology and biochemistry techniques consideredto be familiar to those skilled in the art. Disclosure of suchtechniques can be found, for example, in Sambrook et al., ibid. andAusubel, et al., 1993, Current Protocols in Molecular Biology,Greene/Wiley Interscience, New York, N.Y., and related references.Ausubel, et al, ibid. is incorporated by reference herein in itsentirety. DNA sequence analyses and protein translations were carriedout using the DNAsis program (available from Hitachi Software, SanBruno, Calif.). It should also be noted that since nucleic acidsequencing technology, and in particular the sequencing of PCR products,is not entirely error-free, that the nucleic acid and deduced proteinsequences presented herein represent apparent nucleic acid sequences ofthe nucleic acid molecules encoding D. immitis ankyrin proteins of thepresent invention.

Example 1

This example describes the isolation and sequencing of several D.immitis ankyrin nucleic acid molecules. It is to be noted that some ofthe nucleic acid molecules disclosed in this example were isolated byPCR with degenerate primers and/or primers not fully identical to the D.immitis sequences. The authentic D. immitis nucleotide sequences wereverified on overlapping nucleic acid molecules, and thus the nucleotidesequences of the nucleic acid molecules disclosed in this example arefully representative of D. immitis sequences.

A. As a first step in the isolation of a full-length ankyrin cDNAmolecule from D. immitis, a D. immitis ankyrin nucleic acid molecule ofabout 937 nucleotides, denoted herein as nDiAnk₉₃₇, was isolated from aD. immitis cDNA library by PCR amplification for use as a probe, asfollows. A D. immitis 48-hour L3 cDNA library was constructed in theUni-ZAP® XR vector (available from Stratagene Cloning Systems, La Jolla,Calif.), using Stratagene's ZAP-cDNA® Synthesis Kit protocol and L3mRNAs (i.e., messenger RNAs isolated from D. immitis third-stage larvae,harvested at 48 hours). Initially, degenerate primers were designedbased on conserved regions of the C. elegans UNC-44 gene described inOtsuka, et al., ibid. These initial attempts to PCR-amplify anankyrin-related nucleic acid molecule from the D. immitis cDNA librarywere unsuccessful. In a second attempt, PCR primers were designed basedon the Onchocerca volvulus E1 gene disclosed in Erttmann, et al., 1996a,ibid. These primers included forward primer OVANKY 554+, having thenucleotide sequence 5' CATCAATTTT TGGAATTTTC TGG 3', denoted herein asSEQ ID NO:42 and reverse primer OVANKY 1464-, having the nucleotidesequence 5' CGTTTACAGC AACATCATCC TC 3', denoted herein as SEQ ID NO:43.Several attempts to amplify an ankyrin-related nucleic acid moleculefrom the D. immitis cDNA library with these primers using standard PCRamplification conditions were likewise unsuccessful. Finally, the sameprimers were used in a modified amplification procedure called"touchdown" PCR. This procedure included the following amplificationcycles: six cycles of 94° C. for 30 sec, 58° C. for 45 sec, and 72° C.for 3 min; six cycles of 94° C. for 30 sec, 56° C. for 45 sec, and 72°C. for 3 min; and 20 cycles of 94° C. for 30 sec, 50° C. for 45 sec, and72° C. for 3 min. An about 937-base-pair (bp) DNA fragment was detectedin the PCR reaction by agarose gel electrophoresis. The PCR-amplifiedfragment, denoted herein as nDiAnk₉₃₇, was excised from the gel andpurified using the QIAquick™ kit (available from Qiagen, Chatsworth,Calif.) as per manufacturer's instructions. The resultant 937-bp DNAfragment was subcloned into the pCRII™ vector (available fromInvitrogen, San Diego, Calif.) according to the manufacturer'sinstructions.

The nucleic acid molecule nDiAnk₉₃₇ was sequenced by the Sanger dideoxychain termination method, using the PRISM™ Ready Dye Terminator CycleSequencing Kit with AmpliTaq® DNA Polymerase, FS (available from thePerkin-Elmer Corporation, Norwalk, Conn.). PCR extensions were done inthe GeneAmp™ PCR System 9600 (available from Perkin-Elmer). Excess dyeterminators were removed from extension products using the Centriflex™Gel Filtration Cartridge (available from Advanced Genetics TechnologiesCorporation, Gaithersburg, Md.) following the standard protocol. Sampleswere resuspended according to ABI protocols and were run on aPerkin-Elmer ABI PRISM™ 377 Automated DNA Sequencer. The resultingnucleic acid sequence of nDiAnk₉₃₇ is presented herein as SEQ ID NO:1(the coding strand) and SEQ ID NO:3 (the complementary strand).

Translation of SEQ ID NO:1 yields a protein of about 312 amino acids,denoted herein as PDiAnk₃₁₂, the amino acid sequence of which ispresented in SEQ ID NO:2, assuming a first in-frame codon extending fromnucleotide 1 to nucleotide 3 of SEQ ID NO:1. The coding region encodingPDiAnk₃₁₅ is presented herein as nDiAnk₉₃₆, which has the nucleotidesequence SEQ ID NO:4 (the coding strand) and SEQ ID NO:5 (thecomplementary strand).

B. An additional ankyrin nucleic acid molecule of about 1029 bp,containing the authentic 3' end of a D. immitis ankyrin coding region,was isolated as follows.

Nucleic acid molecule nDiAnk₉₃₇, isolated as disclosed in Example 1A,was used as a hybridization probe to screen a D. immitis 48-hour L3 cDNAlibrary. Nucleic acid molecule nDiAnk₉₃₇ was labeled with α- ³² P! dATPusing the Megaprime™ Kit, available from Amersham, Arlington Heights,Ill. The labeled probe was then hybridized and washed under stringentconditions (i.e., allowing at most about 3% bp mismatch) to about 5×10⁵plaque forming units of an D. immitis 48-hour L3 cDNA library,constructed as disclosed in Example 1A. Fifteen plaques that hybridizedwith the labeled probe were selected and subjected to three rounds ofplaque purification. The size and identity of the bacteriophage clonesthat hybridized with the probe were identified by PCR amplificationusing vector-specific primers, and subsequent Southern hybridization ofthe separated PCR-amplified DNA fragments, using nDiAnk₉₃₇ as a probe.Of these 15 clones, none had inserts larger than about 1.1 kilobases(kb). One positively hybridizing clone having an insert of about 1029 bpwas chosen for further study. The insert from this cDNA clone, denotedherein as nDiAnk₁₀₂₉, was PCR-amplified with a T3/T7 primer set(available from Stratagene), and was subcloned into the pCRII™ vector.The resulting plasmid, containing nucleic acid molecule nDiAnk₁₀₂₉, wassubjected to automated sequencing as described in Example 1A. Thenucleotide sequence of nDiAnk₁₀₂₉ is presented herein as SEQ ID NO:6(the coding strand) and SEQ ID NO:8 (the complementary strand). The 443nucleotides on the 3' end of nDiAnk₉₃₇ lined up with 100% identity tothe 443 nucleotides on the 5' end of nDiANK₁₀₂₉.

Translation of SEQ ID NO:6 yields a protein of about 270 amino acids,denoted herein as PDiAnk₂₇₀, the amino acid sequence of which ispresented in SEQ ID NO:7, assuming a first in-frame codon extending fromnucleotide 2 to nucleotide 4 of SEQ ID NO:6, and a termination codonextending from nucleotide 812 to nucleotide 814 of SEQ ID NO:6. Thecoding region encoding PDiAnk₂₇₀, not including the termination codon,is presented herein as nDiAnk₈₁₀, which has the nucleotide sequence SEQID NO:9 (the coding strand) and SEQ ID NO:10 (the complementary strand).The 147 amino acid residues on the C-terminal end of PDiAnk₃₁₂(disclosed in Example 1A) lined up with 100% identity to the 147 aminoacid residues on the N-terminal end of PDiAnk₂₇₀. PDiAnk₂₇₀ thusrepresents approximately 123 amino acids of new D. immitis ankyrin aminoacid sequence. The presence of a termination codon and a poly-A tail onnDiAnk₁₀₂₉ indicates that nDiAnk₁₀₂₉ represents the authentic 3' end ofthe D. immitis ankyrin messenger RNA that was reverse-transcribed intocDNA.

C. An additional ankyrin nucleic acid molecule of about 600 bp wasisolated by PCR-amplification from a D. immitis 48-hour L3 cDNA library,as follows.

Since nDiAnk₉₃₇ (isolated as disclosed in Example 1A) extended beyondthe 5' end of nDiAnk₁₀₂₉ (isolated as disclosed in Example 1B) by 495bp, nDiAnk₁₀₂₉ did not appear to represent a full-length ankyrin cDNAmolecule. Furthermore, nDiAnk₉₃₇ did not appear to contain the authentic5' end of a full-length ankyrin cDNA molecule. This result suggestedthat the D. immitis ankyrin messenger RNA was larger, but was notrepresented as cDNA at hybridization-detectable levels in the D. immitisL3 cDNA library utilized in these experiments. Therefore, isolation ofadditional portions of a full-length D. immitis ankyrin nucleic acidmolecule were carried out by PCR amplification. An about 600-bp D.immitis ankyrin nucleic acid molecule was amplified by PCR from a D.immitis 48-hour L3 cDNA library, constructed as described in Example 1A,using primers designed according to ankyrin nucleic acid sequences of D.immitis, derived as described in Example 1A, and O. volvulus, derivedfrom the nucleotide sequence reported in Erttmann, et al., 1996a, ibid.The primers included forward primer OVANKY- 1+, having the nucleotidesequence 5' GCACAACCAG TTCCGCAAGA AA 3', denoted herein as SEQ ID NO:44and reverse primer DIANKY- 1-, having the nucleotide sequence 5'GGTTATTGGA AGAAGATTTC C 3', denoted herein as SEQ ID NO:45. DIANKY-1-was designed to hybridize to nucleotides 22-42 of SEQ ID NO:1, i.e.,about 22-42 nucleotides downstream of the 5'-end of nDiAnk₉₃₇.Amplification was accomplished using the "touchdown" PCR protocol asdescribed in Example 1A. A PCR product of about 600 bp was observed uponagarose gel electrophoresis of the PCR reaction, which is denoted hereinas nDiAnk₆₀₀. The PCR product was gel purified and subcloned into thepCRII vector as described in Example 1A. The nucleotide sequence ofnDiAnk₆₀₀ was determined, and is presented herein as SEQ ID NO:11 (thecoding strand) and SEQ ID NO:13 (the complementary strand). The 42nucleotides on the 3' end of nDiAnk₆₀₀ lined up with 100% identity tothe 42 nucleotides on the 5' end of nDiANK₉₃₇ (disclosed in Example 1A).

Translation of SEQ ID NO:11 yields a protein of about 200 amino acids,denoted herein as PDiAnk₂₀₀, the amino acid sequence of which ispresented in SEQ ID NO:12, assuming a first in-frame codon extendingfrom nucleotide 1 to nucleotide 3 of SEQ ID NO:11. The 14 amino acidresidues on the C-terminal end of PDiAnk₂₀₀ lined up with 100% identityto the 14 amino acid residues on the N-terminal end of PDiAnk₃₁₂(disclosed in Example 1A). PDiAnk₂₀₀ thus represents approximately 186amino acids of new D. immitis ankyrin amino acid sequence.

D. An additional D. immitis ankyrin nucleic acid molecule of about 1228bp was isolated from a D. immitis 48-h L3 cDNA library by nested PCR, asfollows.

In order to clone additional D. immitis ankyrin nucleic acid molecules,three degenerate forward primers were designed based on C. elegansUNC-44 DNA sequences reported in Otsuka, et al., ibid. These primerswere paired with various D. immitis reverse primers in severalunsuccessful attempts to isolate additional D. immitis ankyrin nucleicacid molecules from a D. immitis 48-h L3 cDNA library by standard PCRamplification. Finally, an additional D. immitis ankyrin nucleic acidmolecule was isolated when two of the C. elegans-derived primers wereused with two D. immitis-derived primers in a nested PCR. The twoforward primers were: CEANKY-3+, having the nucleotide sequence 5'CAYCARGCNG CNCARCARGG NCA 3', denoted herein as SEQ ID NO:46, andCEANKY-4+, having the nucleotide sequence 5' GTNGAYGAYG TNACNGTNGA YTA3', denoted herein as SEQ ID NO:47. A standard PCR amplification wasperformed using as a template a D. immitis 48-hour L3 cDNA libraryconstructed as described in Example 1A, using forward primer CEANKY-4+and reverse primer DIANKY-1-, as disclosed in Example 1C. No distinctnucleic acid molecules were observed from this PCR reaction upon gelelectrophoresis, but a smear of indistinguishable PCR products wasevident. A very small aliquot of this initial PCR reaction wassubsequently used as a template for a second PCR, using forward primerCEANKY-3+, which was predicted to anneal internal to CEANKY-4+, andreverse primer DIANKY-2-, having the nucleotide sequence 5' GGAATTTGCGACGACGCGGT TC 3', denoted herein as SEQ ID NO:48, which was designed tohybridize to nucleotides 76-97 of SEQ ID NO:11, i.e., about 76-97nucleotides downstream of the 5'-end of nDiAnk₆₀₀. This secondamplification produced a single predominant PCR product of about 1228bp, as viewed by separation on an agarose gel, denoted herein asnDiAnk₁₂₂₈. This PCR product was gel purified and subcloned into plasmidpCRII as described in Example 1A. The nucleotide sequence of nDiAnk₁₂₂₈was determined, and is presented herein as SEQ ID NO:14 (the codingstrand) and SEQ ID NO:16 (the complementary strand). The 97 nucleotideson the 3' end of nDiAnk₁₂₂₈ lined up with 100% identity to the 97nucleotides on the 5' end of nDiANK₆₀₀ (disclosed in Example 1C).

Translation of SEQ ID NO:14 yields a protein of about 409 amino acids,denoted herein as PDiAnk₄₀₉, the amino acid sequence of which ispresented in SEQ ID NO:15, assuming a first in-frame codon extendingfrom nucleotide I to nucleotide 3 of SEQ ID NO:14. The coding regionencoding PDiAnk₄₀₉ is presented herein as nDiAnk₁₂₂₇, which has thenucleotide sequence SEQ ID NO:17 (the coding strand) and SEQ ID NO:18(the complementary strand). The 32 amino acid residues on the C-terminalend of PDiAnk₄₀₉ lined up with 100% identity to the 32 amino acidresidues on the N-terminal end of PDiAnk₂₀₀ (disclosed in Example 1C).PDiAnk₄₀₉ thus represents about 377 amino acids of new D. immitisankyrin amino acid sequence.

E. An additional ankyrin nucleic acid molecule of about 573 bp wasisolated by PCR from a D. immitis 48-hour L3 cDNA library, as follows.

An about 573-bp D. immitis ankyrin nucleic acid molecule was amplifiedby standard PCR from a D. immitis 48-hour L3 cDNA library constructed asdescribed in Example 1A using a primer designed according to ankyrinnucleic acid sequences of D. immitis, derived as described in Example1D, and an M13 reverse primer. The primers included forward primer M13reverse, having the nucleotide sequence 5' CAGGAAACAG CTATGAC 3',denoted herein as SEQ ID NO:49 and reverse primer DIANKY-3-, having thenucleotide sequence 5' TGGAGTTTGT CCTGTCGATG TATG 3', denoted herein asSEQ ID NO:50. DIANKY-3- was designed to hybridize to nucleotides 73-96of SEQ ID NO:14, i.e., about 73-96 nucleotides downstream of the 5'-endof nDiAnk₁₂₂₈. A PCR product of about 573 bp was observed upon agarosegel electrophoresis of the PCR reaction, which is denoted herein asnDiAnk₅₇₃. The PCR product was gel purified and subcloned into the pCRIIvector as described in Example 1A. The nucleotide sequence of nDiAnk₅₇₃was determined, and is presented herein as SEQ ID NO:19 (the codingstrand) and SEQ ID NO:21 (the complementary strand). The 96 nucleotideson the 3' end of nDiAnk₅₇₃ lined up with 100% identity to the 96nucleotides on the 5' end of nDiANK₁₂₂₈ (disclosed in Example 1D).

Translation of SEQ ID NO:19 yields a protein of about 191 amino acids,denoted herein as PDiAnk₁₉₁, the amino acid sequence of which ispresented in SEQ ID NO:20, assuming a first in-frame codon extendingfrom nucleotide 1 to nucleotide 3 of SEQ ID NO:19. The 32 amino acidresidues on the C-terminal end of PDiAnk₁₉₁ lined up with 100% identityto the 32 amino acid residues on the N-terminal end of PDiAnk₄₀₉(disclosed in Example 1D). PDiAnk₁₉₁ thus represents approximately 159amino acids of new D. immitis ankyrin amino acid sequence.

F. An additional ankyrin nucleic acid molecule of about 911 bp wasisolated from first-strand reverse transcriptase cDNA syntheses of D.immitis adult messenger RNA by PCR, as follows.

Attempts to isolate additional portions of a D. immitis ankyrin nucleicacid molecule from a cDNA library were unsuccessful. Therefore,additional portions were isolated from first-strand reversetranscriptase cDNA syntheses of D. immitis adult messenger RNA, by PCRamplification. A D. immitis-specific reverse primer was designed nearthe 5' terminus of nDiAnk₅₇₃ described in Example 1E above. This primer,denoted as DIANKY-4-, having the nucleotide sequence 5' GCTTTGCTTTCAGCATTCGC ATTTGCC 3', denoted herein as SEQ ID NO:51, along withdegenerate forward primer CEANKY-4+, described in Example 1D, were usedin PCR amplifications of first-strand reverse transcriptase cDNAsyntheses of D. immitis adult messenger RNA, prepared by standardmethods. DIANKY-4- was designed to hybridize to nucleotides 138-164 ofSEQ ID NO:19, i.e., about 138-164 nucleotides downstream of the 5'-endof nDiAnk₅₇₃. A PCR product of about 911 bp, visualized by agarose gelelectrophoresis, was obtained by PCR amplification using as a templatefirst strand cDNA syntheses of either adult female or adult male D.immitis messenger RNA using these primers. It should be noted that PCRamplifications performed with three other degenerate forward primers,designed from the UNC-44 nucleotide sequence of Otsuka, et al., ibid.,in conjunction with D. immitis reverse primers, did not result in thesuccessful amplification of D. immitis ankyrin nucleic acid molecules.The 911-bp PCR products were excised from the agarose gel on which theywere separated, and bathed in a small volume of Tris-EDTA buffer. Tenmicroliter aliquots of these excised products in TE were used astemplate for reamplification PCRs, using the same primers, to verify theproduct. More intense bands of exactly the same size, denoted herein asnDiAnk₉₁₁, were produced by these second round PCRs, as seen byseparation on an agarose gel.

Nucleic acid molecules from both the first and second PCR reactions weregel purified and subcloned into plasmid pCRII as described in Example1A. The subcloned DNA products were submitted for automated sequencing.Sequence analysis revealed that the inserts of all PCR products wereidentical, and are denoted herein as SEQ ID NO:22 (the coding strand)and SEQ ID NO:24 (the complementary strand). The 164 nucleotides on the3' end of nDiAnk₉₁₁, lined up with 100% identity to the 164 nucleotideson the 5' end of nDiANK₅₇₃ (disclosed in Example 1E).

Translation of SEQ ID NO:22 yields a protein of about 303 amino acids,denoted herein as PDiAnk₃₀₃, the amino acid sequence of which ispresented in SEQ ID NO:23, assuming a first in-frame codon extendingfrom nucleotide 1 to nucleotide 3 of SEQ ID NO:22. The coding regionencoding PDiAnk₃₀₃ is presented herein as nDiAnk₉₀₉, which has thenucleotide sequence SEQ ID NO:25 (the coding strand) and SEQ ID NO:26(the complementary strand). The 54 amino acid residues on the C-terminalend of PDiAnk₃₀₃ lined up with 100% identity to the 54 amino acidresidues on the N-terminal end of the PDiAnk₁₉₁ (disclosed in Example1E). PDiAnk₃₀₃ thus represents approximately 249 amino acids of new D.immitis ankyrin amino acid sequence.

G. An additional ankyrin nucleic acid molecule of about 1096 bp,containing sequences representing the authentic 5' end of a D. immitisankyrin messenger RNA, was isolated from first-strand reversetranscriptase cDNA syntheses of D. immitis adult messenger RNA by PCR,as follows.

A D. immitis-specific reverse primer was designed near the 5' terminusof nDiAnk₉₁₁ described in Example 1F above. This primer, denotedDIANKY-7-, having the nucleotide sequence 5' GTGAGATAGT CAACAGTAACATCATCC 3', denoted herein as SEQ ID NO:53, was designed to hybridize tonucleotides 3-29 of SEQ ID NO:22, i.e., about 3-29 nucleotidesdownstream of the 5'-end of nDiAnk ₉₁₁. DIANKY-7- was used along with asense primer designed according to the nematode splice leader (SL) inPCR amplifications of first-strand reverse transcriptase cDNA synthesesof D. immitis adult messenger RNA, prepared by standard methods. Most,but not all nematode messenger RNAs have the SL at their 5' ends, andthe presence of the 5' SL sequence is indicative of an apparently fulllength cDNA molecule. See, for example Blaxter and Liu, 1996, Int. J.Parasitol. 26, 1025-1033, which is incorporated herein by reference. Thesplice leader primer, denoted DiSL, has the nucleotide sequence 5'GGTTTAATTA CCCAAGTTTG AG 3', denoted herein as SEQ ID NO:52. Using theseprimers, PCR products of about 1096 bp were obtained using adult maleand adult female mRNAs as templates. These nucleic acid molecules weregel-purified using the QIAquick™ kit as per manufacturer's instructions.The yield of these purifications was low, so one microliter each of thepurified DNA products were used as templates in reamplification PCRsusing the same primers. More intense products of precisely the samesize, collectively denoted herein as nDiANK₁₀₉₆, were obtained from thereamplification PCRs, and were subcloned into plasmid pCRII as describedin Example 1A. One of the subcloned nucleic acid molecules was submittedfor automated nucleic acid sequencing. Sequence analysis suggested thatnDiAnk₁₀₉₆, the sequence of which is represented herein as SEQ ID NO:27(the coding strand) and SEQ ID NO:29 (the complementary strand),represented the authentic 5' end of a D. immitis ankyrin messenger RNA.The 5' end of nDiAnk₁₀₉₆ included the spliced leader sequence, 28nucleotides of 5' untranslated sequence, and the starting methionine ofthe coding sequence. The 29 nucleotides on the 3' end of nDiAnk₁₀₉₆lined up with 100% identity to the 29 nucleotides on the 5' end ofnDiANK₉₁₁ (disclosed in Example 1F).

Translation of SEQ ID NO:27 yields a protein of about 348 amino acids,denoted herein as PDiAnk₃₄₈, the amino acid sequence of which ispresented in SEQ ID NO:28, assuming a start codon extending fromnucleotide 51 to nucleotide 53 of SEQ ID NO:27. The nucleic acidmolecule representing the coding region encoding PDiAnk₃₄₈, denotedherein as nDiAnk₁₀₄₄, is presented herein as SEQ ID NO:30 (the codingstrand) and SEQ ID NO:31 (the complementary strand). The about 9 aminoacid residues on the C-terminal end of PDiAnk₃₄₈ lined up with about100% identity to the about 9 amino acid residues on the N-terminal endof the PDiAnk₃₀₃ (disclosed in Example 1F). PDiAnk₃₄₈ thus representsapproximately 339 amino acids of new D. immitis ankyrin amino acidsequence.

H. A composite nucleotide sequence of a full-length D. immitis ankyringene was compiled as follows.

A D. immitis ankyrin nucleic acid molecule of 5503 bp, denoted herein asnDiAnk₅₅₀₃, including an apparently full-length coding region, wascompiled by aligning the overlapping nucleic acid sequences SEQ ID NO:1,SEQ ID NO:6, SEQ ID NO:11, SEQ ID NO:14, SEQ ID NO:19, SEQ ID NO:22, andSEQ ID NO:27. The composite full-length nucleic acid molecule has anucleic acid sequence presented herein as SEQ ID NO:32 (the codingstrand) and SEQ ID NO:34 (the complementary strand). SEQ ID NO:32contains the 5' nematode spliced leader sequence extending from aboutnucleotide 1 through about nucleotide 27, and a 28 bp 5' non-codingregion extending from about nucleotide 28 through about nucleotide 50.

Translation of SEQ ID NO:32 yields a full-length protein of about 1745amino acids, denoted PDiAnk₁₇₄₅, assuming a start codon extending fromnucleotide 51 through nucleotide 53 of SEQ ID NO:32, and a stop codonextending from nucleotide 5286 through nucleotide 5285 of SEQ ID NO:32.The resulting amino acid sequence is presented as SEQ ID NO:33. Thecoding region encoding PDiAnk₁₇₄₅, not including the termination codonis denoted herein as nDiANK₅₂₃₅, and has the nucleotide sequence SEQ IDNO:35 (the coding strand) and SEQ ID NO:36 (the complementary strand).SEQ ID NO:35 is predicted to encode a protein with a molecular mass ofabout 191.7 kD and with a predicted pI of about 5.76, as calculated bythe DNAsis program.

A homology search of a non-redundant protein database was performed withSEQ ID NO:33, using the blastp program available through the BLAST™network of the National Center for Biotechnology Information (NCBI)(National Library of Medicine, National Institutes of Health, Baltimore,Md.), available on the World Wide Web. This database includesSwissProt+PIR+SPupdate+GenPept+GPUpdate+PDB databases. The highestscoring match of the homology search at the amino acid level wasGenBank™ accession number gil1208874, a C. elegans ankyrin-like protein,to which SEQ ID NO:33 showed about 69% identity, spanning from aboutamino acid 1 through about amino acid 1745 of SEQ ID NO:33. The secondhighest highest scoring match of the homology search at the amino acidlevel was GenBank™ accession number gil406288, a human brain ankyrinprotein (variant I, Ankyrin_(B)), to which SEQ ID NO:33 showed about 51%identity, spanning from about amino acid 1 through about amino acid1745. SEQ ID NO:33 was also compared to the sequence of the O. volvulusE1 protein as disclosed by Erttmann, et al., 1996a, ibid. A region ofSEQ ID NO:33 spanning from about amino acid 1282 to about amino acid1745 showed about 78% identity to the full-length O. volvulus E1 proteinof 467 amino acids. At the nucleotide level, the coding regionrepresented in SEQ ID NO:35 was compared to the cDNA encoding the O.volvulus E1 protein. A region of SEQ ID NO:35 spanning from aboutnucleotide 3423 to about nucleotide 5574 showed about 88% nucleic acididentity to the cDNA containing a full-length coding region of 1401nucleotides encoding the O. volvulus E1 protein.

Example 2

This Example discloses an analysis of the predicted functional domainsof a full-length D. immitis ankyrin protein.

Based on comparisons with mammalian ankyrin proteins, as well as theprotein encoded by the C. elegans UNC-44 gene, the putative positions ofthe three functional domains of ankyrin proteins were identified in SEQID NO:33, the amino acid sequence of an apparent full-length D. immitisankyrin protein, namely PDiAnk₁₇₄₅, isolated as disclosed in Example 1.While not being bound by theory, an N-terminal membrane protein-bindingdomain is predicted to extend from about amino acid 1 to about aminoacid 880 of SEQ ID NO:33, a spectrin-binding domain is predicted toextend from about amino acid 881 to about amino acid 1398 of SEQ IDNO:33, and a regulatory domain is predicted to extend from about aminoacid 1399 to about amino acid 1745 of SEQ ID NO:33.

The N-terminal membrane protein-binding domain of PDiAnk₁₇₄₅ is furthercharacterized by the presence of 24 tandemly arrayed repeats, most ofwhich comprise about 33 amino acid residues. These repeat regions arelisted in Table 1, designated by numbers, along with a general consensussequence pattern. The terms "start" and "end" in Table 1 refer to thesequential amino acid numbers in SEQ ID NO:33 corresponding to the firstand last amino acid, respectively, of each repeat listed in Table 1. Thefirst and the twenty-fourth repeats are not as well conserved as theother repeats. To the inventor's knowledge, this is the first disclosureof ankyrin-like repeats in a parasitic nematode.

                                      TABLE 1                                     __________________________________________________________________________    Ankyrin-like repeats of PDiAnk.sub.1745                                       No.                                                                              START                          END                                         __________________________________________________________________________    01 36  ESSASFLRAA                                                                            RAGNLDRVLE                                                                            LLRSGTDINT                                                                            CNA                                                                               68                                         02 69  NGLNALHLAS                                                                            KEGHHEVVRE                                                                            LLKRKADVDA                                                                            ATR                                                                              101                                         03 102 KGNTALHIAS                                                                            LAGQELIVTV                                                                            LVENGAVVNV                                                                            QSL                                                                              134                                         04 135 NGFTPLYMAA                                                                            QENHESVVRY                                                                            LLAHNANQAL                                                                            STE                                                                              167                                         05 168 DGFTPLAVAL                                                                            QQGHDRVVAV                                                                            LLENDTRGK˜                                                                      ˜˜˜                                                            196                                         06 197 VRLPALHIAA                                                                            KKDDTKAATL                                                                            LLQNEHNSDV                                                                            TSK                                                                              229                                         07 230 SGFTPLHIAA                                                                            HYGNENVAQL                                                                            LLEKGANVNY                                                                            QAR                                                                              262                                         08 263 HNISPLHVAT                                                                            KWGRTNMVSL                                                                            LLAHGAVIDC                                                                            RTR                                                                              295                                         09 296 DLLTPLHCAS                                                                            RSGHDQVVDL                                                                            LLEKGAPISA                                                                            KTK                                                                              328                                         10 329 NGLAPLHMAA                                                                            Q˜˜˜˜˜˜˜˜.a                   bout.   ˜˜˜˜˜˜˜VD                           D       VTV                                                                              345                                         11 346 DYLTPLHVAA                                                                            HCGHVRVAKL                                                                            LLDRNADPNA                                                                            RAL                                                                              378                                         12 379 NGFTPLHIAC                                                                            KKNRIKIVEL                                                                            LLKYHAAIEA                                                                            TTE                                                                              411                                         13 412 SGLSPLHVAA                                                                            FMGAINIVIY                                                                            LLQQGANADV                                                                            ATV                                                                              444                                         14 445 RGETPLHLAA                                                                            RANQTDIVRV                                                                            LVRNGAQVDA                                                                            AAR                                                                              477                                         15 478 ELQTPLHIAS                                                                            RLGNTDIVIL                                                                            LLQANASPNA                                                                            ATR                                                                              510                                         16 511 DLYTPLHIAA                                                                            KEGQEEVAAI                                                                            LMDHGTDKTL                                                                            LTK                                                                              543                                         17 544 KGFTPLHLAA                                                                            KYGNLPVAKS                                                                            LLERGTPVDI                                                                            EGK                                                                              576                                         18 577 NQVTPLHVAA                                                                            HYNNDKVALL                                                                            LLENGASAHA                                                                            AAK                                                                              609                                         19 610 NGYTPLHIAA                                                                            KKNQMDIAST                                                                            LLHYKANANA                                                                            ESK                                                                              642                                         20 643 AGFTPLHLAA                                                                            QEGHREMAAL                                                                            LIENGAKVGA                                                                            QAR                                                                              675                                         21 676 NGLTPMHLCA                                                                            QEDRVSVAEE                                                                            LVKENAAIDP                                                                            KTK                                                                              708                                         22 709 AGYTPLHVAC                                                                            HFGQINMVRF                                                                            LIEHGARVSV                                                                            ITR                                                                              741                                         23 742 ASYTPLHQAA                                                                            QQGHNSVVRY                                                                            LLEHGASPNV                                                                            HTS                                                                              774                                         24 775 TGQTPLSIAE                                                                            RLGYVSVVEA                                                                            LKTITETTVI                                                                            TET                                                                              807                                         Consensus                                                                            -G-TPLH-AA                                                                            --GH---V--                                                                            LL--GA--N-                                                                            ---                                                           A       D                                                      __________________________________________________________________________

Repeat regions 01 through 24 as disclosed in Table 1 are presentedherein as SEQ ID NO:61 through SEQ ID NO:84, respectively. The consensussequence is presented herein as SEQ ID NO:85. A novel feature of themembrane-binding domain of PDiAnk₁₇₄₅ is repeat No. 10 (SEQ ID NO:70),which apparently comprises only 17 amino acids. While not being bound bytheory, this shortened repeat is unique among ankyrin proteins.

Example 3

This Example demonstrates the use of D. immitis nucleic acid moleculesof the present invention to obtain, by PCR amplification, an ankyrinnucleic acid molecule from a related filariid nematode, Brugia malayi.

A B. malayi ankyrin nucleic acid molecule was PCR amplified from afirst-strand cDNA synthesis of messenger RNA prepared from B. malayiadult female worms, as follows. The PCR primers included forward primerCEANKY 4+, as disclosed in Example 1D, and reverse primer DIANKY 4-, asdisclosed in Example 1F. An about 908-bp nucleic acid molecule wasamplified in a PCR amplification of a first-strand reverse transcriptasecDNA synthesis of B. malayi adult female messenger RNA, prepared bystandard methods, and is denoted herein as nBmAnk₉₀₈.

Nucleic acid molecule nBmAnk₉₀₈ was gel purified and subcloned intoplasmid pCRII, and sequenced as described in Example 1A. The sequence ispresented as SEQ ID NO:37 (the coding strand) and SEQ ID NO:39 (thecomplementary strand). Translation of SEQ ID NO:37 yields anon-full-length protein of about 302 amino acids, denoted herein asPBmAnk₃₀₂, assuming a first in-frame codon extending from nucleotide 1through nucleotide 3 of SEQ ID NO:37. The resulting amino acid sequenceis presented as SEQ ID NO:38. The coding region encoding PBmAnk₃₀₂ isdenoted herein as nBmANK₉₀₆, and has the nucleotide sequence SEQ IDNO:40 (the coding strand) and SEQ ID NO:41 (the complementary strand).

A homology search of a non-redundant protein database was performed onSEQ ID NO:38 using the BLAST network. The homology spans from aboutamino acid 1 through amino acid 302 of SEQ ID NO:38. The highest scoringmatch of the homology search at the amino acid level was GenBankaccession number A57282, a C. elegans ankyrin-like protein, which wasabout 86% identical to SEQ ID NO:38 through a region extending fromabout amino acid 353 through about amino acid 654 of A57282. The B.malayi ankyrin nucleic acid molecule and protein sequences representedby SEQ ID NO:37 and SEQ ID NO:38, respectively, had no similarity to theO. volvulus E1 nucleic acid molecule and protein disclosed by Erttmann,et al., 1996a, ibid.

The amino acid sequence of SEQ ID NO:38 was also compared to PDiAnk₁₇₄₅(i.e., SEQ ID NO:33 of the present invention). PBmAnk₃₀₂ had 95%identity to the region of SEQ ID NO:33 spanning from about amino acid341 through about amino acid 642.

Example 4

The following experiment was performed in order to confirm the origin ofthe ankyrin gene in the D. immitis genome, and to identify genomicrestriction fragments associating with a partial ankyrin cDNA clone. Theexperiment also evaluates if multiple copies of the ankyrin gene arepresent in the D. immitis genome. Four enzymes were each usedindividually to digest about 10 micrograms of D. immitis genomic DNAeach. A Southern blot containing genomic DNA samples restricted withSau3A-I, EcoRI, HindIII, and XhoI, respectively, was hybridized understringent conditions using nDiAnk₉₃₇, isolated as disclosed in Example1A, labeled with the radioactive isotope ³² P. The nDiAnk₉₃₇ probehybridized to three bands of 460 bp, 550 bp, and 770 bp in the genomicrestriction digestion using Sau3A-I. A single band of 5490 bp hybridizedin the EcoRI digestion. Two bands of 930 bp and 3330 bp hybridized inthe HindIII digestion. Two bands also hybridized in the XhoI digestion(which of the four enzymes digested the genomic DNA most incompletely),one of 3330 bp and one of 43,200 bp, the latter appearing in a bandrepresenting undigested DNA. This experiment confirmed the D. immitisorigin of ankyrin nucleic acid molecules of the present invention. Whilenot being bound by theory, since no EcoRI sites are present within thefull-length coding region of nDiAnk₁₇₄₅, the single EcoRI genomicfragment hybridizing with the nDiAnk₉₃₇ probe suggests that at least theportion of nDiAnk₁₇₄₅ included in nDiAnk₉₃₇ is present as a single copyin the D. immitis genome.

Example 5

This Example discloses the production of a recombinant cell of thepresent invention.

Recombinant molecule pTrc-nDiAnk₁₈₆₆, containing a D. immitis ankyrinnucleic acid molecule operatively linked to trc transcription controlsequences and to a fusion sequence encoding the T7 tag and apoly-histidine segment, was produced in the following manner. An about1866-nucleotide DNA fragment containing nucleotides spanning from about3423 through about 5288 of SEQ ID NO:32, denoted herein as nDiAnk₁₈₆₆,was PCR-amplified by "touchdown" PCR from D. immitis L3 and L4 cDNAlibraries produced by the methods described in Example 1A, using senseprimer OVANKY 1+(SEQ ID NO:44, as described in Example 1C), and DIANKY-,having the nucleotide sequence 5' CCGGAATTCT TATTCATGAA CGCTTTGCCC TTT3', denoted herein as SEQ ID NO:55, EcoRI site in bold. DIANKY- wasdesigned to anneal to a region of SEQ ID NO:32 extending from nucleotide5365 through nucleotide 5288. This PCR product was gel-purified usingthe QIAquick™ kit as per manufacturer's instructions. The yield of thispurification was low, so one microliter of the purified DNA product wasused as template in a reamplification PCR (standard PCR) using forwardprimer DIANKY 3+, having the nucleotide sequence 5' CGCGGATCCGGCACAACCAG TTCCGCAAGA A 3', denoted herein as SEQ ID NO:54, BamHI sitein bold, and antisense primer DIANKY-, as described above. DIANKY 3+ wasdesigned to anneal to the PCR product generated above, but shares 17/20bp in common with nucleotides 3423 through 3443 of SEQ ID NO:32. A moreintense product of approximately the same size, denoted herein asnDiAnk₁₈₆₆, was obtained from the reamplification PCR. Recombinantmolecule pTrc-nDiAnk₁₈₆₆ was produced by digesting the PCR-amplified DNAfragment with BamHI and EcoRI restriction endonucleases, gel purifyingthe resulting fragment and directionally subcloning it into expressionvector pTrcHisB (available from Invitrogen) that had been cleaved withBamHI and EcoRI and gel purified.

Recombinant molecule pTrc-nDiAnk₈₆₆ was transformed into E. coli to formrecombinant cell E. coli:pTrc-nDiAnk₁₈₆₆ using standard techniques.

Example 6

This Example discloses the production of additional recombinant cells ofthe present invention.

A. Recombinant molecule pTrc-nDiAnk₂₃₆₁, containing a D. immitis ankyrinnucleic acid molecule encoding the predicted membrane protein-bindingdomain (disclosed in Example 2), operatively linked to trc transcriptioncontrol sequences and to a fusion sequence encoding the T7 tag and apoly-histidine segment, is produced in the following manner. An about2361-nucleotide DNA fragment spanning from about nucleotide 51 throughabout nucleotide 2411 of SEQ ID NO:32, denoted herein as nDiAnk₂₃₆₁, wasRT-PCR-amplified from four different D. immitis messenger RNA templates.The templates were as follows: 48-hour L3 (i.e., third-stage larvaeharvested at 48 hours), 6-day L4 (i.e., fourth stage larvae harvested at6 days), adult female, and adult male D. immitis mRNAs. The PCR primersincluded DIANKY REP+, having the nucleotide sequence 5'CGCGGATCCGATGAGTAATC CTATAGTCGA GGG 3', denoted herein as SEQ ID NO:56, BamHI sitein bold, and reverse primer DIANKY REP-,having the nucleotide sequence5' CCGGAATTCC GGTTACCCTA GACGTTCAGC AATCG 3', denoted herein as SEQ IDNO:57, EcoRI site in bold. The amplification produced a product of thepredicted size from each of the four mRNA templates. This resultindicates that the portion of the D. immitis ankyrin gene encoding theputative membrane protein-binding domain is expressed in at least twolarval stages and in male and female adult parasites. Recombinantmolecule pTrc-nDiAnk₂₃₆₁ is produced by digesting any one of thePCR-amplified DNA fragments with BamHI and EcoRI restrictionendonucleases, gel purifying the resulting fragment and directionallysubcloning it into expression vector pTrcHisB (available fromInvitrogen) which is cleaved with BamHI and EcoRI and gel purified.

B. Recombinant molecule pTrc-nDiAnk₁₂₆₆, containing a D. immitis ankyrinnucleic acid molecule encoding the predicted spectrin-binding domain(disclosed in Example 2), operatively linked to trc transcriptioncontrol sequences and to a fusion sequence encoding the T7 tag and apoly-histidine segment, is produced in the following manner. An about1266-nucleotide DNA fragment spanning from about nucleotide 2850 throughabout nucleotide 4115 of SEQ ID NO:32, denoted herein as nDiAnk₁₂₆₆, wasRT-PCR-amplified from the four different D. immitis messenger RNAtemplates described in Example 6A above. The PCR primers included DIANKYSPECT B+, having the nucleotide sequence 5' CCGGGATCCG CGCGCACGTGGAGGAGCAAT GCGT 3', denoted herein as SEQ ID NO:58, BamHI site in bold,and reverse primer DIANKY SPECT B-, having the nucleotide sequence 5'CGCGAATTCC GGTTATTCGT TGTCCGTGTG AGTGCG 3', denoted herein as SEQ IDNO:59, EcoRI site in bold. The amplification produced a product of thepredicted size from each of the four mRNA templates. This resultindicates that the portion of the D. immitis ankyrin gene encoding theputative spectrin-binding domain is expressed in at least two larvalstages and in male and female adult parasites. Recombinant moleculepTrc-nDiAnk₁₂₆₆ is produced by digesting any one of the PCR-amplifiedDNA fragments with BamHI and EcoRI restriction endonucleases, gelpurifying the resulting fragment and directionally subcloning it intoexpression vector pTrcHisB (available from Invitrogen) which is cleavedwith BamHI and EcoRI and gel purified.

C. Recombinant molecule pTrc-nDiAnk₈₆₄, containing a D. immitis ankyrinnucleic acid molecule encoding the predicted regulatory domain(disclosed in Example 2), operatively linked to trc transcriptioncontrol sequences and to a fusion sequence encoding the T7 tag and apoly-histidine segment, is produced in the following manner. An about864-nucleotide DNA fragment spanning from about nucleotide 4425 throughabout nucleotide 5288 of SEQ ID NO:32, denoted herein as nDiAnk₈₆₄, wasRT-PCR-amplified from the four different D. immitis messenger RNAtemplates described in Example 6A above. The PCR primers included DIANKYREG+, having the nucleotide sequence 5' CGCGGATCCG CGCCAACTAG TTGGTCTTGAAGCAGTC 3', denoted herein as SEQ ID NO:60, BamHI site in bold, andreverse primer DIANKY-, having the nucleotide sequence 5' CCGGAATTCTTATTCATGAA CGCTTTGCCC TTT 3', denoted herein as SEQ ID NO:55, EcoRI sitein bold. The amplification produced a product of the predicted size fromeach of the four mRNA templates. This result indicates that the portionof the D. immitis ankyrin gene encoding the putative regulatory domainis expressed in at least two larval stages and in male and female adultparasites. Recombinant molecule pTrc-nDiAnk₈₆₄ is produced by digestingthe PCR-amplified DNA fragment with BamHI and EcoRI restrictionendonucleases, gel purifying the resulting fragment and directionallysubcloning it into expression vector pTrcHisB (available fromInvitrogen) which is cleaved with BamHI and EcoRI and gel purified.

Recombinant molecules pTrc-nDiAnk₂₃₆₁, pTrc-nDiAnk₁₂₆₆, andpTrc-nDiAnk₈₆₄ are transformed into E. coli to form recombinant cells E.coli:pTrc-nDiAnk₂₃₆₁, E. coli:pTrc-nDiAnk₁₂₆₆, and E.coli:pTrc-nDiAnk₈₆₄ using standard techniques.

Example 7

This example demonstrates the production of D. immitis ankyrin proteinsof the present invention in prokaryotic cells.

Recombinant cell E. coli:pTrc-nDiAnk₁₈₆₆, produced as described inExample 5, was cultured in shake-flasks containing an enriched bacterialgrowth medium containing 0.1 mg/ml ampicillin at about 37° C. When thecells reached an OD₆₀₀ of about 0.5, expression of D. immitis nucleicacid molecule nDiAnk₁₈₆₆ was induced by addition of about 0.5 mM IPTG,and the cells were cultured for about 3 hr at about 37° C. Proteinproduction was monitored by SDS-polyacrylamide gel electrophoresis(SDS-PAGE) of recombinant cell lysates, followed by Coomassie bluestaining, using standard techniques. Recombinant cell E.coli:pTrc-nDiAnk₁₈₆₆ produced a fusion protein, denoted herein asPHIS-PDiANK₆₂₂, that migrated with an apparent molecular weight of about98 kD, although the predicted molecular weight is only about 74 kD.While not being bound by theory, the difference between the observed andpredicted molecular weights may be attributed to the known acidity ofthe regulatory domains of ankyrin proteins, which is known by thoseskilled in the art to reduce the binding of SDS to the protein, therebyresulting in an aberrant migration in SDS-PAGE.

Immunoblot analysis of recombinant cell E. coli:pTrc-nDiAnk₁₈₆₆ lysatesindicated that the about 98-kD protein was able to bind to a T7 tagmonoclonal antibody (available from Novagen, Inc., Madison, Wis.)directed against the fusion portion of the recombinant PHIS-PDiANK₆₂₂fusion protein.

Recombinant cells E. coli:pTrc-nDiAnk₂₃₆₁, E. coli:pTrc-nDiAnk₁₂₆₆, andE. coli:pTrc-nDiAnk₈₆₄, produced as described in Example 6, are culturedseparately in shake-flasks containing an enriched bacterial growthmedium containing 0.1 mg/ml ampicillin at about 37° C. When the cellsreach an OD₆₀₀ of about 0.5, expression of D. immitis nucleic acidmolecules nDiAnk₂₃₆₁, nDiAnk₁₂₆₆, and nDiAnk₈₆₄, respectively, isinduced by addition of about 0.5 mM IPTG, and the cells are cultured forabout 3 hr at about 37° C. Protein production is monitored bySDS-polyacrylamide gel electrophoresis (SDS-PAGE) of recombinant celllysates, followed by Coomassie blue staining, using standard techniques.Recombinant cells E. coli:pTrc-nDiAnk₂₃₆₁, E. coli:pTrc-nDiAnk₁₂₆₆, andE. coli:pTrc-nDiAnk₈₆₄ are expected to produce a fusion proteins,denoted herein as PHIS-PDiANK₇₈₇, PHIS-PDiANK₄₃₃, and PHIS-PDiANK₂₈₈,respectively.

SEQUENCE LISTING

The following Sequence Listing is submitted pursuant to 37 CFR § 1.821.A copy in computer readable form is also submitted herewith.

Applicants assert pursuant to 37 CFR § 1.821 (f) that the content of thepaper and computer readable copies of SEQ ID NO:1 through SEQ ID NO:85submitted herewith are the same.

While various embodiments of the present invention have been describedin detail, it is apparent that modifications and adaptations of thoseembodiments will occur to those skilled in the art. It is to beexpressly understood, however, that such modifications and adaptationsare within the scope of the present invention, as set forth in thefollowing claims.

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 85                                                 (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 937 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..936                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       CATCAGTTTTTGGAATTTTCTGGAAATCTTCTTCCAATAACC42                                  HisGlnPheLeuGluPheSerGlyAsnLeuLeuProIleThr                                    1510                                                                          AAGAGTGGTGACCAACTTTCTCTTTATTTTCTACCATTCCAA84                                  LysSerGlyAspGlnLeuSerLeuTyrPheLeuProPheGln                                    152025                                                                        GAAAATCGTCTTGCTTTCATGGTAAAGATACGCACTCACACG126                                 GluAsnArgLeuAlaPheMetValLysIleArgThrHisThr                                    303540                                                                        GACAACGAAACTGCAGCTGATGGCCGGATAGTATTTATGAAA168                                 AspAsnGluThrAlaAlaAspGlyArgIleValPheMetLys                                    455055                                                                        GAACCAAAATTGAGAGCCGAAAATTTACCTCCGCAGACGCCA210                                 GluProLysLeuArgAlaGluAsnLeuProProGlnThrPro                                    606570                                                                        GTGTGTACTCTTGCAATCACTCTTCCGGAATACACTGGGCCG252                                 ValCysThrLeuAlaIleThrLeuProGluTyrThrGlyPro                                    7580                                                                          GAGCCGATGGTTTCCAAAAAACTCTTCTATTCGGAAGCTTCT294                                 GluProMetValSerLysLysLeuPheTyrSerGluAlaSer                                    859095                                                                        TTGACTGAGAAATACGTTGGAGCTTTCCATGAAACTGCTGAA336                                 LeuThrGluLysTyrValGlyAlaPheHisGluThrAlaGlu                                    100105110                                                                     CCTGATAACTTGCCACTAGCACATGTTGCACTATTAATTGGC378                                 ProAspAsnLeuProLeuAlaHisValAlaLeuLeuIleGly                                    115120125                                                                     GCTGATTGGCATCGGTTAGCTCGAGCGCTTGAAGTACCTGAT420                                 AlaAspTrpHisArgLeuAlaArgAlaLeuGluValProAsp                                    130135140                                                                     ATTGATATACGACAAGTTCGACATCAACTAGTTGGTCTTGAA462                                 IleAspIleArgGlnValArgHisGlnLeuValGlyLeuGlu                                    145150                                                                        GCAGTCACTATTCTACGTATTTGGATATTTTTGAAGAAAGAA504                                 AlaValThrIleLeuArgIleTrpIlePheLeuLysLysGlu                                    155160165                                                                     CAAGCTACGCCCGTTGCTTTGCGATCAGCATTGCAGCGAATA546                                 GlnAlaThrProValAlaLeuArgSerAlaLeuGlnArgIle                                    170175180                                                                     GGACGTGATGATGTTGTACGAGAAATGGATCGAGCTGAAAAG588                                 GlyArgAspAspValValArgGluMetAspArgAlaGluLys                                    185190195                                                                     CTAGATGGTTTAGAAGGAACACCTGTATCGCATATTTCTGGA630                                 LeuAspGlyLeuGluGlyThrProValSerHisIleSerGly                                    200205210                                                                     CCCTCAATAACTCTGTCATCTACTTTGCTAGAGGTAGCAGGC672                                 ProSerIleThrLeuSerSerThrLeuLeuGluValAlaGly                                    215220                                                                        GACAGACGTCGTCACGCCGAGGTAACAATGGCGCAACAGCGA714                                 AspArgArgArgHisAlaGluValThrMetAlaGlnGlnArg                                    225230235                                                                     TTGGCACAAGAACCGTTTTTTCAGCAAGTAGGGTATAATGGG756                                 LeuAlaGlnGluProPhePheGlnGlnValGlyTyrAsnGly                                    240245250                                                                     ACACCTGGAGATCCAGAAGAACCCAAAGAACAGTCATTCCAC798                                 ThrProGlyAspProGluGluProLysGluGlnSerPheHis                                    255260265                                                                     GAAGAGGAAGAGGAAGTTGCAGTTTCAGAAATTCGAACAGTT840                                 GluGluGluGluGluValAlaValSerGluIleArgThrVal                                    270275280                                                                     GTGCGCACTGAACGACATGTGCATGATTCGGAAAATGGTCCT882                                 ValArgThrGluArgHisValHisAspSerGluAsnGlyPro                                    285290                                                                        ATTGTGGAAGAGCGTACAATAACAACTACGTATGAGGATGAT924                                 IleValGluGluArgThrIleThrThrThrTyrGluAspAsp                                    295300305                                                                     GTTGCTGTAAACG937                                                              ValAlaValAsn                                                                  310                                                                           (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 312 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       HisGlnPheLeuGluPheSerGlyAsnLeuLeuProIleThr                                    1510                                                                          LysSerGlyAspGlnLeuSerLeuTyrPheLeuProPheGln                                    152025                                                                        GluAsnArgLeuAlaPheMetValLysIleArgThrHisThr                                    303540                                                                        AspAsnGluThrAlaAlaAspGlyArgIleValPheMetLys                                    455055                                                                        GluProLysLeuArgAlaGluAsnLeuProProGlnThrPro                                    606570                                                                        ValCysThrLeuAlaIleThrLeuProGluTyrThrGlyPro                                    7580                                                                          GluProMetValSerLysLysLeuPheTyrSerGluAlaSer                                    859095                                                                        LeuThrGluLysTyrValGlyAlaPheHisGluThrAlaGlu                                    100105110                                                                     ProAspAsnLeuProLeuAlaHisValAlaLeuLeuIleGly                                    115120125                                                                     AlaAspTrpHisArgLeuAlaArgAlaLeuGluValProAsp                                    130135140                                                                     IleAspIleArgGlnValArgHisGlnLeuValGlyLeuGlu                                    145150                                                                        AlaValThrIleLeuArgIleTrpIlePheLeuLysLysGlu                                    155160165                                                                     GlnAlaThrProValAlaLeuArgSerAlaLeuGlnArgIle                                    170175180                                                                     GlyArgAspAspValValArgGluMetAspArgAlaGluLys                                    185190195                                                                     LeuAspGlyLeuGluGlyThrProValSerHisIleSerGly                                    200205210                                                                     ProSerIleThrLeuSerSerThrLeuLeuGluValAlaGly                                    215220                                                                        AspArgArgArgHisAlaGluValThrMetAlaGlnGlnArg                                    225230235                                                                     LeuAlaGlnGluProPhePheGlnGlnValGlyTyrAsnGly                                    240245250                                                                     ThrProGlyAspProGluGluProLysGluGlnSerPheHis                                    255260265                                                                     GluGluGluGluGluValAlaValSerGluIleArgThrVal                                    270275280                                                                     ValArgThrGluArgHisValHisAspSerGluAsnGlyPro                                    285290                                                                        IleValGluGluArgThrIleThrThrThrTyrGluAspAsp                                    295300305                                                                     ValAlaValAsn                                                                  310                                                                           (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 937 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       CGTTTACAGCAACATCATCCTCATACGTAGTTGTTATTGTACGCTCTTCC50                          ACAATAGGACCATTTTCCGAATCATGCACATGTCGTTCAGTGCGCACAAC100                         TGTTCGAATTTCTGAAACTGCAACTTCCTCTTCCTCTTCGTGGAATGACT150                         GTTCTTTGGGTTCTTCTGGATCTCCAGGTGTCCCATTATACCCTACTTGC200                         TGAAAAAACGGTTCTTGTGCCAATCGCTGTTGCGCCATTGTTACCTCGGC250                         GTGACGACGTCTGTCGCCTGCTACCTCTAGCAAAGTAGATGACAGAGTTA300                         TTGAGGGTCCAGAAATATGCGATACAGGTGTTCCTTCTAAACCATCTAGC350                         TTTTCAGCTCGATCCATTTCTCGTACAACATCATCACGTCCTATTCGCTG400                         CAATGCTGATCGCAAAGCAACGGGCGTAGCTTGTTCTTTCTTCAAAAATA450                         TCCAAATACGTAGAATAGTGACTGCTTCAAGACCAACTAGTTGATGTCGA500                         ACTTGTCGTATATCAATATCAGGTACTTCAAGCGCTCGAGCTAACCGATG550                         CCAATCAGCGCCAATTAATAGTGCAACATGTGCTAGTGGCAAGTTATCAG600                         GTTCAGCAGTTTCATGGAAAGCTCCAACGTATTTCTCAGTCAAAGAAGCT650                         TCCGAATAGAAGAGTTTTTTGGAAACCATCGGCTCCGGCCCAGTGTATTC700                         CGGAAGAGTGATTGCAAGAGTACACACTGGCGTCTGCGGAGGTAAATTTT750                         CGGCTCTCAATTTTGGTTCTTTCATAAATACTATCCGGCCATCAGCTGCA800                         GTTTCGTTGTCCGTGTGAGTGCGTATCTTTACCATGAAAGCAAGACGATT850                         TTCTTGGAATGGTAGAAAATAAAGAGAAAGTTGGTCACCACTCTTGGTTA900                         TTGGAAGAAGATTTCCAGAAAATTCCAAAAACTGATG937                                      (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 936 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       CATCAGTTTTTGGAATTTTCTGGAAATCTTCTTCCAATAACCAAGAGTGG50                          TGACCAACTTTCTCTTTATTTTCTACCATTCCAAGAAAATCGTCTTGCTT100                         TCATGGTAAAGATACGCACTCACACGGACAACGAAACTGCAGCTGATGGC150                         CGGATAGTATTTATGAAAGAACCAAAATTGAGAGCCGAAAATTTACCTCC200                         GCAGACGCCAGTGTGTACTCTTGCAATCACTCTTCCGGAATACACTGGGC250                         CGGAGCCGATGGTTTCCAAAAAACTCTTCTATTCGGAAGCTTCTTTGACT300                         GAGAAATACGTTGGAGCTTTCCATGAAACTGCTGAACCTGATAACTTGCC350                         ACTAGCACATGTTGCACTATTAATTGGCGCTGATTGGCATCGGTTAGCTC400                         GAGCGCTTGAAGTACCTGATATTGATATACGACAAGTTCGACATCAACTA450                         GTTGGTCTTGAAGCAGTCACTATTCTACGTATTTGGATATTTTTGAAGAA500                         AGAACAAGCTACGCCCGTTGCTTTGCGATCAGCATTGCAGCGAATAGGAC550                         GTGATGATGTTGTACGAGAAATGGATCGAGCTGAAAAGCTAGATGGTTTA600                         GAAGGAACACCTGTATCGCATATTTCTGGACCCTCAATAACTCTGTCATC650                         TACTTTGCTAGAGGTAGCAGGCGACAGACGTCGTCACGCCGAGGTAACAA700                         TGGCGCAACAGCGATTGGCACAAGAACCGTTTTTTCAGCAAGTAGGGTAT750                         AATGGGACACCTGGAGATCCAGAAGAACCCAAAGAACAGTCATTCCACGA800                         AGAGGAAGAGGAAGTTGCAGTTTCAGAAATTCGAACAGTTGTGCGCACTG850                         AACGACATGTGCATGATTCGGAAAATGGTCCTATTGTGGAAGAGCGTACA900                         ATAACAACTACGTATGAGGATGATGTTGCTGTAAAC936                                       (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 936 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       GTTTACAGCAACATCATCCTCATACGTAGTTGTTATTGTACGCTCTTCCA50                          CAATAGGACCATTTTCCGAATCATGCACATGTCGTTCAGTGCGCACAACT100                         GTTCGAATTTCTGAAACTGCAACTTCCTCTTCCTCTTCGTGGAATGACTG150                         TTCTTTGGGTTCTTCTGGATCTCCAGGTGTCCCATTATACCCTACTTGCT200                         GAAAAAACGGTTCTTGTGCCAATCGCTGTTGCGCCATTGTTACCTCGGCG250                         TGACGACGTCTGTCGCCTGCTACCTCTAGCAAAGTAGATGACAGAGTTAT300                         TGAGGGTCCAGAAATATGCGATACAGGTGTTCCTTCTAAACCATCTAGCT350                         TTTCAGCTCGATCCATTTCTCGTACAACATCATCACGTCCTATTCGCTGC400                         AATGCTGATCGCAAAGCAACGGGCGTAGCTTGTTCTTTCTTCAAAAATAT450                         CCAAATACGTAGAATAGTGACTGCTTCAAGACCAACTAGTTGATGTCGAA500                         CTTGTCGTATATCAATATCAGGTACTTCAAGCGCTCGAGCTAACCGATGC550                         CAATCAGCGCCAATTAATAGTGCAACATGTGCTAGTGGCAAGTTATCAGG600                         TTCAGCAGTTTCATGGAAAGCTCCAACGTATTTCTCAGTCAAAGAAGCTT650                         CCGAATAGAAGAGTTTTTTGGAAACCATCGGCTCCGGCCCAGTGTATTCC700                         GGAAGAGTGATTGCAAGAGTACACACTGGCGTCTGCGGAGGTAAATTTTC750                         GGCTCTCAATTTTGGTTCTTTCATAAATACTATCCGGCCATCAGCTGCAG800                         TTTCGTTGTCCGTGTGAGTGCGTATCTTTACCATGAAAGCAAGACGATTT850                         TCTTGGAATGGTAGAAAATAAAGAGAAAGTTGGTCACCACTCTTGGTTAT900                         TGGAAGAAGATTTCCAGAAAATTCCAAAAACTGATG936                                       (2) INFORMATION FOR SEQ ID NO:6:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1029 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 2..811                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       GAAGAAAGAACAAGCTACGCCCGTTGCTTTGCGATCAGCA40                                    LysLysGluGlnAlaThrProValAlaLeuArgSerAla                                       1510                                                                          TTGCAGCGAATAGGACGTGATGATGTTGTACGAGAAATGGAT82                                  LeuGlnArgIleGlyArgAspAspValValArgGluMetAsp                                    152025                                                                        CGAGCTGAAAAGCTAGATGGTTTAGAAGGAACACCTGTATCG124                                 ArgAlaGluLysLeuAspGlyLeuGluGlyThrProValSer                                    303540                                                                        CATATTTCTGGACCCTCAATAACTCTGTCATCTACTTTGCTA166                                 HisIleSerGlyProSerIleThrLeuSerSerThrLeuLeu                                    455055                                                                        GAGGTAGCAGGCGACAGACGTCGTCACGCCGAGGTAACAATG208                                 GluValAlaGlyAspArgArgArgHisAlaGluValThrMet                                    6065                                                                          GCGCAACAGCGATTGGCACAAGAACCGTTTTTTCAGCAAGTA250                                 AlaGlnGlnArgLeuAlaGlnGluProPhePheGlnGlnVal                                    707580                                                                        GGGTATAATGGGACACCTGGAGATCCAGAAGAACCCAAAGAA292                                 GlyTyrAsnGlyThrProGlyAspProGluGluProLysGlu                                    859095                                                                        CAGTCATTCCACGAAGAGGAAGAGGAAGTTGCAGTTTCAGAA334                                 GlnSerPheHisGluGluGluGluGluValAlaValSerGlu                                    100105110                                                                     ATTCGAACAGTTGTGCGCACTGAACGACATGTGCATGATTCG376                                 IleArgThrValValArgThrGluArgHisValHisAspSer                                    115120125                                                                     GAAAATGGTCCTATTGTGGAAGAGCGTACAATAACAACTACG418                                 GluAsnGlyProIleValGluGluArgThrIleThrThrThr                                    130135                                                                        TATGAGGATGATGTTGCTGTAAACGAAGAAGAAATTGTTGAC460                                 TyrGluAspAspValAlaValAsnGluGluGluIleValAsp                                    140145150                                                                     AAAATAGTGCCTCTCAACGAAGAGGAGCAAGAAAAATGGGAT502                                 LysIleValProLeuAsnGluGluGluGlnGluLysTrpAsp                                    155160165                                                                     CGAATGGTTCGAGAAGTGGAAATGAATTTTGAGCAACAAGAA544                                 ArgMetValArgGluValGluMetAsnPheGluGlnGlnGlu                                    170175180                                                                     ACATCAAAAGAAGGAACGTTTGGTTGTCAGACAACACATGAG586                                 ThrSerLysGluGlyThrPheGlyCysGlnThrThrHisGlu                                    185190195                                                                     AAAGAAAAAGATGATGATGGTGGCAGTCTGAAGACGACAATG628                                 LysGluLysAspAspAspGlyGlySerLeuLysThrThrMet                                    200205                                                                        AAAGATAGTCACGTAAGGCAGATTTTCTTCGATGGAGGTGAG670                                 LysAspSerHisValArgGlnIlePhePheAspGlyGlyGlu                                    210215220                                                                     ACATCCGCTAATGAAACAGGATTAAGTAGCGGGGATGCAGAC712                                 ThrSerAlaAsnGluThrGlyLeuSerSerGlyAspAlaAsp                                    225230235                                                                     ACTATTATGACTCCAACGACAAAGGAGGATAATCATGTTATA754                                 ThrIleMetThrProThrThrLysGluAspAsnHisValIle                                    240245250                                                                     GACGTAATGGAGGAAAGGCGAACTGATGAAGAGGCCAAAGGG796                                 AspValMetGluGluArgArgThrAspGluGluAlaLysGly                                    255260265                                                                     CAAAGCGTTCATGAATAATCTGGATCCACAAATTGATTTAAATCGCAA844                           GlnSerValHisGlu                                                               270                                                                           TCTCGCACATGCCTATGTTGCTAATATTTAATGAAATTTTTCAAAGCAAT894                         AATTTGAATGCTGTTTGGGCTTCCCATATTGTTAAAGCGTTTTCCATCGT944                         CCATTCACTTTTTGTTTTTGCTGTAGTCTGTAACTGCTACTCTTGATAAA994                         TTTGCTCCAGTAAAAAAAAAAAAAAAAAAAAAAAA1029                                       (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 270 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       LysLysGluGlnAlaThrProValAlaLeuArgSerAlaLeu                                    1510                                                                          GlnArgIleGlyArgAspAspValValArgGluMetAspArg                                    152025                                                                        AlaGluLysLeuAspGlyLeuGluGlyThrProValSerHis                                    303540                                                                        IleSerGlyProSerIleThrLeuSerSerThrLeuLeuGlu                                    455055                                                                        ValAlaGlyAspArgArgArgHisAlaGluValThrMetAla                                    606570                                                                        GlnGlnArgLeuAlaGlnGluProPhePheGlnGlnValGly                                    7580                                                                          TyrAsnGlyThrProGlyAspProGluGluProLysGluGln                                    859095                                                                        SerPheHisGluGluGluGluGluValAlaValSerGluIle                                    100105110                                                                     ArgThrValValArgThrGluArgHisValHisAspSerGlu                                    115120125                                                                     AsnGlyProIleValGluGluArgThrIleThrThrThrTyr                                    130135140                                                                     GluAspAspValAlaValAsnGluGluGluIleValAspLys                                    145150                                                                        IleValProLeuAsnGluGluGluGlnGluLysTrpAspArg                                    155150165                                                                     MetValArgGluValGluMetAsnPheGluGlnGlnGluThr                                    170175180                                                                     SerLysGluGlyThrPheGlyCysGlnThrThrHisGluLys                                    185190195                                                                     GluLysAspAspAspGlyGlySerLeuLysThrThrMetLys                                    200205210                                                                     AspSerHisValArgGlnIlePhePheAspGlyGlyGluThr                                    215220                                                                        SerAlaAsnGluThrGlyLeuSerSerGlyAspAlaAspThr                                    225230235                                                                     IleMetThrProThrThrLysGluAspAsnHisValIleAsp                                    240245250                                                                     ValMetGluGluArgArgThrAspGluGluAlaLysGlyGln                                    255260265                                                                     SerValHisGlu                                                                  270                                                                           (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1029 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       TTTTTTTTTTTTTTTTTTTTTTTTACTGGAGCAAATTTATCAAGAGTAGC50                          AGTTACAGACTACAGCAAAAACAAAAAGTGAATGGACGATGGAAAACGCT100                         TTAACAATATGGGAAGCCCAAACAGCATTCAAATTATTGCTTTGAAAAAT150                         TTCATTAAATATTAGCAACATAGGCATGTGCGAGATTGCGATTTAAATCA200                         ATTTGTGGATCCAGATTATTCATGAACGCTTTGCCCTTTGGCCTCTTCAT250                         CAGTTCGCCTTTCCTCCATTACGTCTATAACATGATTATCCTCCTTTGTC300                         GTTGGAGTCATAATAGTGTCTGCATCCCCGCTACTTAATCCTGTTTCATT350                         AGCGGATGTCTCACCTCCATCGAAGAAAATCTGCCTTACGTGACTATCTT400                         TCATTGTCGTCTTCAGACTGCCACCATCATCATCTTTTTCTTTCTCATGT450                         GTTGTCTGACAACCAAACGTTCCTTCTTTTGATGTTTCTTGTTGCTCAAA500                         ATTCATTTCCACTTCTCGAACCATTCGATCCCATTTTTCTTGCTCCTCTT550                         CGTTGAGAGGCACTATTTTGTCAACAATTTCTTCTTCGTTTACAGCAACA600                         TCATCCTCATACGTAGTTGTTATTGTACGCTCTTCCACAATAGGACCATT650                         TTCCGAATCATGCACATGTCGTTCAGTGCGCACAACTGTTCGAATTTCTG700                         AAACTGCAACTTCCTCTTCCTCTTCGTGGAATGACTGTTCTTTGGGTTCT750                         TCTGGATCTCCAGGTGTCCCATTATACCCTACTTGCTGAAAAAACGGTTC800                         TTGTGCCAATCGCTGTTGCGCCATTGTTACCTCGGCGTGACGACGTCTGT850                         CGCCTGCTACCTCTAGCAAAGTAGATGACAGAGTTATTGAGGGTCCAGAA900                         ATATGCGATACAGGTGTTCCTTCTAAACCATCTAGCTTTTCAGCTCGATC950                         CATTTCTCGTACAACATCATCACGTCCTATTCGCTGCAATGCTGATCGCA1000                        AAGCAACGGGCGTAGCTTGTTCTTTCTTC1029                                             (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 810 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       AAGAAAGAACAAGCTACGCCCGTTGCTTTGCGATCAGCATTGCAGCGAAT50                          AGGACGTGATGATGTTGTACGAGAAATGGATCGAGCTGAAAAGCTAGATG100                         GTTTAGAAGGAACACCTGTATCGCATATTTCTGGACCCTCAATAACTCTG150                         TCATCTACTTTGCTAGAGGTAGCAGGCGACAGACGTCGTCACGCCGAGGT200                         AACAATGGCGCAACAGCGATTGGCACAAGAACCGTTTTTTCAGCAAGTAG250                         GGTATAATGGGACACCTGGAGATCCAGAAGAACCCAAAGAACAGTCATTC300                         CACGAAGAGGAAGAGGAAGTTGCAGTTTCAGAAATTCGAACAGTTGTGCG350                         CACTGAACGACATGTGCATGATTCGGAAAATGGTCCTATTGTGGAAGAGC400                         GTACAATAACAACTACGTATGAGGATGATGTTGCTGTAAACGAAGAAGAA450                         ATTGTTGACAAAATAGTGCCTCTCAACGAAGAGGAGCAAGAAAAATGGGA500                         TCGAATGGTTCGAGAAGTGGAAATGAATTTTGAGCAACAAGAAACATCAA550                         AAGAAGGAACGTTTGGTTGTCAGACAACACATGAGAAAGAAAAAGATGAT600                         GATGGTGGCAGTCTGAAGACGACAATGAAAGATAGTCACGTAAGGCAGAT650                         TTTCTTCGATGGAGGTGAGACATCCGCTAATGAAACAGGATTAAGTAGCG700                         GGGATGCAGACACTATTATGACTCCAACGACAAAGGAGGATAATCATGTT750                         ATAGACGTAATGGAGGAAAGGCGAACTGATGAAGAGGCCAAAGGGCAAAG800                         CGTTCATGAA810                                                                 (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 810 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      TTCATGAACGCTTTGCCCTTTGGCCTCTTCATCAGTTCGCCTTTCCTCCA50                          TTACGTCTATAACATGATTATCCTCCTTTGTCGTTGGAGTCATAATAGTG100                         TCTGCATCCCCGCTACTTAATCCTGTTTCATTAGCGGATGTCTCACCTCC150                         ATCGAAGAAAATCTGCCTTACGTGACTATCTTTCATTGTCGTCTTCAGAC200                         TGCCACCATCATCATCTTTTTCTTTCTCATGTGTTGTCTGACAACCAAAC250                         GTTCCTTCTTTTGATGTTTCTTGTTGCTCAAAATTCATTTCCACTTCTCG300                         AACCATTCGATCCCATTTTTCTTGCTCCTCTTCGTTGAGAGGCACTATTT350                         TGTCAACAATTTCTTCTTCGTTTACAGCAACATCATCCTCATACGTAGTT400                         GTTATTGTACGCTCTTCCACAATAGGACCATTTTCCGAATCATGCACATG450                         TCGTTCAGTGCGCACAACTGTTCGAATTTCTGAAACTGCAACTTCCTCTT500                         CCTCTTCGTGGAATGACTGTTCTTTGGGTTCTTCTGGATCTCCAGGTGTC550                         CCATTATACCCTACTTGCTGAAAAAACGGTTCTTGTGCCAATCGCTGTTG600                         CGCCATTGTTACCTCGGCGTGACGACGTCTGTCGCCTGCTACCTCTAGCA650                         AAGTAGATGACAGAGTTATTGAGGGTCCAGAAATATGCGATACAGGTGTT700                         CCTTCTAAACCATCTAGCTTTTCAGCTCGATCCATTTCTCGTACAACATC750                         ATCACGTCCTATTCGCTGCAATGCTGATCGCAAAGCAACGGGCGTAGCTT800                         GTTCTTTCTT810                                                                 (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 600 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..600                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      GCCCAGCCAGTTCCACAAGAGATAGTCACTCGTTTACATGGG42                                  AlaGlnProValProGlnGluIleValThrArgLeuHisGly                                    1510                                                                          AATAGAGTCGCTGTTTCTCCAATTGTAACTGTTGAACCGCGT84                                  AsnArgValAlaValSerProIleValThrValGluProArg                                    152025                                                                        CGTCGCAAATTCCATAAGCCCATAACGCTGTGCATACCATTG126                                 ArgArgLysPheHisLysProIleThrLeuCysIleProLeu                                    303540                                                                        CCACAAAGCTCAAATAAAGGAATGTTAACACAATATAGTGGC168                                 ProGlnSerSerAsnLysGlyMetLeuThrGlnTyrSerGly                                    455055                                                                        CAACCAGGACAGGAACCACCGACGCTGCGTTTACTCTGCAGT210                                 GlnProGlyGlnGluProProThrLeuArgLeuLeuCysSer                                    606570                                                                        AAAACTGGAGGTTCTTCTCCTGCACAGTGGGAAGATATTACT252                                 LysThrGlyGlySerSerProAlaGlnTrpGluAspIleThr                                    7580                                                                          GGAACTACCCAGTTAACATTTACTGGTGAGGACGTTTCATTT294                                 GlyThrThrGlnLeuThrPheThrGlyGluAspValSerPhe                                    859095                                                                        ACAACTACGGTTTCTGCTCGATTTTGGTTGATGGATTGCCAA336                                 ThrThrThrValSerAlaArgPheTrpLeuMetAspCysGln                                    100105110                                                                     ACTCCGCGAGATGCGGCACGAATGGCACAAGAAGTTTACAAT378                                 ThrProArgAspAlaAlaArgMetAlaGlnGluValTyrAsn                                    115120125                                                                     GAAGCAATTGCAGTTCCTTATATGGCTAAATTTCTTATTTTT420                                 GluAlaIleAlaValProTyrMetAlaLysPheLeuIlePhe                                    130135140                                                                     GCTCGACGAACTTTTCCTGCCGAAGGACAGTTGAGATTGTTT462                                 AlaArgArgThrPheProAlaGluGlyGlnLeuArgLeuPhe                                    145150                                                                        TGTATGACTGATGATCGGGAAGATAAAACCCTGGAAAAACAA504                                 CysMetThrAspAspArgGluAspLysThrLeuGluLysGln                                    155160165                                                                     GAACGTTTCATTGAAATTGCGAAATCGAAAGATGTAGAAGTC546                                 GluArgPheIleGluIleAlaLysSerLysAspValGluVal                                    170175180                                                                     TTAAGTGGGCGACATCAGTTTTTGGAATTTTCTGGAAATCTT588                                 LeuSerGlyArgHisGlnPheLeuGluPheSerGlyAsnLeu                                    185190195                                                                     CTTCCAATAACC600                                                               LeuProIleThr                                                                  200                                                                           (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 200 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      AlaGlnProValProGlnGluIleValThrArgLeuHisGly                                    1510                                                                          AsnArgValAlaValSerProIleValThrValGluProArg                                    152025                                                                        ArgArgLysPheHisLysProIleThrLeuCysIleProLeu                                    303540                                                                        ProGlnSerSerAsnLysGlyMetLeuThrGlnTyrSerGly                                    455055                                                                        GlnProGlyGlnGluProProThrLeuArgLeuLeuCysSer                                    606570                                                                        LysThrGlyGlySerSerProAlaGlnTrpGluAspIleThr                                    7580                                                                          GlyThrThrGlnLeuThrPheThrGlyGluAspValSerPhe                                    859095                                                                        ThrThrThrValSerAlaArgPheTrpLeuMetAspCysGln                                    100105110                                                                     ThrProArgAspAlaAlaArgMetAlaGlnGluValTyrAsn                                    115120125                                                                     GluAlaIleAlaValProTyrMetAlaLysPheLeuIlePhe                                    130135140                                                                     AlaArgArgThrPheProAlaGluGlyGlnLeuArgLeuPhe                                    145150                                                                        CysMetThrAspAspArgGluAspLysThrLeuGluLysGln                                    155160165                                                                     GluArgPheIleGluIleAlaLysSerLysAspValGluVal                                    170175180                                                                     LeuSerGlyArgHisGlnPheLeuGluPheSerGlyAsnLeu                                    185190195                                                                     LeuProIleThr                                                                  200                                                                           (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 600 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      GGTTATTGGAAGAAGATTTCCAGAAAATTCCAAAAACTGATGTCGCCCAC50                          TTAAGACTTCTACATCTTTCGATTTCGCAATTTCAATGAAACGTTCTTGT100                         TTTTCCAGGGTTTTATCTTCCCGATCATCAGTCATACAAAACAATCTCAA150                         CTGTCCTTCGGCAGGAAAAGTTCGTCGAGCAAAAATAAGAAATTTAGCCA200                         TATAAGGAACTGCAATTGCTTCATTGTAAACTTCTTGTGCCATTCGTGCC250                         GCATCTCGCGGAGTTTGGCAATCCATCAACCAAAATCGAGCAGAAACCGT300                         AGTTGTAAATGAAACGTCCTCACCAGTAAATGTTAACTGGGTAGTTCCAG350                         TAATATCTTCCCACTGTGCAGGAGAAGAACCTCCAGTTTTACTGCAGAGT400                         AAACGCAGCGTCGGTGGTTCCTGTCCTGGTTGGCCACTATATTGTGTTAA450                         CATTCCTTTATTTGAGCTTTGTGGCAATGGTATGCACAGCGTTATGGGCT500                         TATGGAATTTGCGACGACGCGGTTCAACAGTTACAATTGGAGAAACAGCG550                         ACTCTATTCCCATGTAAACGAGTGACTATCTCTTGTGGAACTGGCTGGGC600                         (2) INFORMATION FOR SEQ ID NO:14:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1228 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..1227                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      CATCAAGCTGCTCAGCAAGGGCATAACAGTGTTGTACGTTAC42                                  HisGlnAlaAlaGlnGlnGlyHisAsnSerValValArgTyr                                    1510                                                                          TTGTTGGAACATGGTGCAAGTCCAAATGTTCATACATCGACA84                                  LeuLeuGluHisGlyAlaSerProAsnValHisThrSerThr                                    152025                                                                        GGACAAACTCCATTATCGATTGCTGAACGTCTAGGGTATGTA126                                 GlyGlnThrProLeuSerIleAlaGluArgLeuGlyTyrVal                                    303540                                                                        TCCGTGGTTGAAGCGCTTAAAACAATTACCGAGACTACTGTG168                                 SerValValGluAlaLeuLysThrIleThrGluThrThrVal                                    455055                                                                        ATAACGGAGACCACAACCGTTACTGAAGAAAGATATAAACCT210                                 IleThrGluThrThrThrValThrGluGluArgTyrLysPro                                    606570                                                                        CAGAATCCCGAAGCAATGAATGAAACCATGTTTTCCGATTCC252                                 GlnAsnProGluAlaMetAsnGluThrMetPheSerAspSer                                    7580                                                                          GAAGATGAAGGTGAAGATAATCAGATCACAGCCAATGCTCAT294                                 GluAspGluGlyGluAspAsnGlnIleThrAlaAsnAlaHis                                    859095                                                                        GCTCATGATTTCTCAGAAAGCCTCACAAAAGGTTTGCACGAT336                                 AlaHisAspPheSerGluSerLeuThrLysGlyLeuHisAsp                                    100105110                                                                     TCAACTGGTGTACATTTGATTCATGCCACAGAACCGACATTG378                                 SerThrGlyValHisLeuIleHisAlaThrGluProThrLeu                                    115120125                                                                     TCACGAAGTCCGGAAGTGGAAGGTACGGATGGCGATTTGGAT420                                 SerArgSerProGluValGluGlyThrAspGlyAspLeuAsp                                    130135140                                                                     GCCTTAATTCGTAAAGCACAACATGAACCAATTACTACAGCG462                                 AlaLeuIleArgLysAlaGlnHisGluProIleThrThrAla                                    145150                                                                        ATGGCCGATCCTTCCTTAGATGCATCGCTTCCTGACAATGTT504                                 MetAlaAspProSerLeuAspAlaSerLeuProAspAsnVal                                    155160165                                                                     ACGATAATGAGAACTACCATGCAACCTAGTTTTTTAATTTCG546                                 ThrIleMetArgThrThrMetGlnProSerPheLeuIleSer                                    170175180                                                                     TTTATGGTGGATGCACGTGGAGGAGCAATGCGTGGTTGTAGG588                                 PheMetValAspAlaArgGlyGlyAlaMetArgGlyCysArg                                    185190195                                                                     CATTCCGGTGTCAGAATCATTATACCACCGAGGAAAGCGCCG630                                 HisSerGlyValArgIleIleIleProProArgLysAlaPro                                    200205210                                                                     CAACCTACACGGGTCACATGCAGATACCTTGGAAAGGACAAG672                                 GlnProThrArgValThrCysArgTyrLeuGlyLysAspLys                                    215220                                                                        TTAGCGCATCCACCACCATTAAGTGAAGGTGAAGCGCTCGCN714                                 LeuAlaHisProProProLeuSerGluGlyGluAlaLeuAla                                    225230235                                                                     TCACGTATACTTGAAATGGCACCACATGGAGCAAAATTCTTA756                                 SerArgIleLeuGluMetAlaProHisGlyAlaLysPheLeu                                    240245250                                                                     GGCCCTGTTATATTGGAAGTACCACATTTTGCATCACTTCGT798                                 GlyProValIleLeuGluValProHisPheAlaSerLeuArg                                    255260265                                                                     GGACGAGAGAGAGAGATTGTCATTTTGCGTTCTGATGATGGG840                                 GlyArgGluArgGluIleValIleLeuArgSerAspAspGly                                    270275280                                                                     CAGCATTGGAAAGAGCATCAGCTTGAAGCAACAGAAGATGCT882                                 GlnHisTrpLysGluHisGlnLeuGluAlaThrGluAspAla                                    285290                                                                        GTACAAGAGGTGCTCAATGAATCGTTTGATGCAGAAGAGTTG924                                 ValGlnGluValLeuAsnGluSerPheAspAlaGluGluLeu                                    295300305                                                                     TCGCAACTTGATGATTTGCATACATCACGGATTACGCGTATC966                                 SerGlnLeuAspAspLeuHisThrSerArgIleThrArgIle                                    310315320                                                                     CTGACCAATGATTTCCCAATGTATTTCGCGGTCGTTACTCGT1008                                LeuThrAsnAspPheProMetTyrPheAlaValValThrArg                                    325330335                                                                     GTGCGGCAAGAAGTGCACTGTGTTGGTCCAGAAGGTGGTGTA1050                                ValArgGlnGluValHisCysValGlyProGluGlyGlyVal                                    340345350                                                                     ATACTCTCTTCAGTTGTTCCTCATGTGCAGGCCATATTTCCG1092                                IleLeuSerSerValValProHisValGlnAlaIlePhePro                                    355360                                                                        GATGGTTCCTTGACTAAGACGATCAAAGTATCTGTGCAAGCC1134                                AspGlySerLeuThrLysThrIleLysValSerValGlnAla                                    365370375                                                                     CAGCCAGTTCCACAAGAGATAGTCACTCGTTTACATGGGAAT1176                                GlnProValProGlnGluIleValThrArgLeuHisGlyAsn                                    380385390                                                                     AGAGTCGCTGTTTCTCCAATTGTAACTGTTGAACCGCGTCGT1218                                ArgValAlaValSerProIleValThrValGluProArgArg                                    395400405                                                                     CGCAAATTCC1228                                                                ArgLysPhe                                                                     (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 409 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                      HisGlnAlaAlaGlnGlnGlyHisAsnSerValValArgTyr                                    1510                                                                          LeuLeuGluHisGlyAlaSerProAsnValHisThrSerThr                                    152025                                                                        GlyGlnThrProLeuSerIleAlaGluArgLeuGlyTyrVal                                    303540                                                                        SerValValGluAlaLeuLysThrIleThrGluThrThrVal                                    455055                                                                        IleThrGluThrThrThrValThrGluGluArgTyrLysPro                                    606570                                                                        GlnAsnProGluAlaMetAsnGluThrMetPheSerAspSer                                    7580                                                                          GluAspGluGlyGluAspAsnGlnIleThrAlaAsnAlaHis                                    859095                                                                        AlaHisAspPheSerGluSerLeuThrLysGlyLeuHisAsp                                    100105110                                                                     SerThrGlyValHisLeuIleHisAlaThrGluProThrLeu                                    115120125                                                                     SerArgSerProGluValGluGlyThrAspGlyAspLeuAsp                                    130135140                                                                     AlaLeuIleArgLysAlaGlnHisGluProIleThrThrAla                                    145150                                                                        MetAlaAspProSerLeuAspAlaSerLeuProAspAsnVal                                    155160165                                                                     ThrIleMetArgThrThrMetGlnProSerPheLeuIleSer                                    170175180                                                                     PheMetValAspAlaArgGlyGlyAlaMetArgGlyCysArg                                    185190195                                                                     HisSerGlyValArgIleIleIleProProArgLysAlaPro                                    200205210                                                                     GlnProThrArgValThrCysArgTyrLeuGlyLysAspLys                                    215220                                                                        LeuAlaHisProProProLeuSerGluGlyGluAlaLeuAla                                    225230235                                                                     SerArgIleLeuGluMetAlaProHisGlyAlaLysPheLeu                                    240245250                                                                     GlyProValIleLeuGluValProHisPheAlaSerLeuArg                                    255260265                                                                     GlyArgGluArgGluIleValIleLeuArgSerAspAspGly                                    270275280                                                                     GlnHisTrpLysGluHisGlnLeuGluAlaThrGluAspAla                                    285290                                                                        ValGlnGluValLeuAsnGluSerPheAspAlaGluGluLeu                                    295300305                                                                     SerGlnLeuAspAspLeuHisThrSerArgIleThrArgIle                                    310315320                                                                     LeuThrAsnAspPheProMetTyrPheAlaValValThrArg                                    325330335                                                                     ValArgGlnGluValHisCysValGlyProGluGlyGlyVal                                    340345350                                                                     IleLeuSerSerValValProHisValGlnAlaIlePhePro                                    355360                                                                        AspGlySerLeuThrLysThrIleLysValSerValGlnAla                                    365370375                                                                     GlnProValProGlnGluIleValThrArgLeuHisGlyAsn                                    380385390                                                                     ArgValAlaValSerProIleValThrValGluProArgArg                                    395400405                                                                     ArgLysPhe                                                                     (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1228 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      GGAATTTGCGACGACGCGGTTCAACAGTTACAATTGGAGAAACAGCGACT50                          CTATTCCCATGTAAACGAGTGACTATCTCTTGTGGAACTGGCTGGGCTTG100                         CACAGATACTTTGATCGTCTTAGTCAAGGAACCATCCGGAAATATGGCCT150                         GCACATGAGGAACAACTGAAGAGAGTATTACACCACCTTCTGGACCAACA200                         CAGTGCACTTCTTGCCGCACACGAGTAACGACCGCGAAATACATTGGGAA250                         ATCATTGGTCAGGATACGCGTAATCCGTGATGTATGCAAATCATCAAGTT300                         GCGACAACTCTTCTGCATCAAACGATTCATTGAGCACCTCTTGTACAGCA350                         TCTTCTGTTGCTTCAAGCTGATGCTCTTTCCAATGCTGCCCATCATCAGA400                         ACGCAAAATGACAATCTCTCTCTCTCGTCCACGAAGTGATGCAAAATGTG450                         GTACTTCCAATATAACAGGGCCTAAGAATTTTGCTCCATGTGGTGCCATT500                         TCAAGTATACGTGANGCGAGCGCTTCACCTTCACTTAATGGTGGTGGATG550                         CGCTAACTTGTCCTTTCCAAGGTATCTGCATGTGACCCGTGTAGGTTGCG600                         GCGCTTTCCTCGGTGGTATAATGATTCTGACACCGGAATGCCTACAACCA650                         CGCATTGCTCCTCCACGTGCATCCACCATAAACGAAATTAAAAAACTAGG700                         TTGCATGGTAGTTCTCATTATCGTAACATTGTCAGGAAGCGATGCATCTA750                         AGGAAGGATCGGCCATCGCTGTAGTAATTGGTTCATGTTGTGCTTTACGA800                         ATTAAGGCATCCAAATCGCCATCCGTACCTTCCACTTCCGGACTTCGTGA850                         CAATGTCGGTTCTGTGGCATGAATCAAATGTACACCAGTTGAATCGTGCA900                         AACCTTTTGTGAGGCTTTCTGAGAAATCATGAGCATGAGCATTGGCTGTG950                         ATCTGATTATCTTCACCTTCATCTTCGGAATCGGAAAACATGGTTTCATT1000                        CATTGCTTCGGGATTCTGAGGTTTATATCTTTCTTCAGTAACGGTTGTGG1050                        TCTCCGTTATCACAGTAGTCTCGGTAATTGTTTTAAGCGCTTCAACCACG1100                        GATACATACCCTAGACGTTCAGCAATCGATAATGGAGTTTGTCCTGTCGA1150                        TGTATGAACATTTGGACTTGCACCATGTTCCAACAAGTAACGTACAACAC1200                        TGTTATGCCCTTGCTGAGCAGCTTGATG1228                                              (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1227 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      CATCAAGCTGCTCAGCAAGGGCATAACAGTGTTGTACGTTACTTGTTGGA50                          ACATGGTGCAAGTCCAAATGTTCATACATCGACAGGACAAACTCCATTAT100                         CGATTGCTGAACGTCTAGGGTATGTATCCGTGGTTGAAGCGCTTAAAACA150                         ATTACCGAGACTACTGTGATAACGGAGACCACAACCGTTACTGAAGAAAG200                         ATATAAACCTCAGAATCCCGAAGCAATGAATGAAACCATGTTTTCCGATT250                         CCGAAGATGAAGGTGAAGATAATCAGATCACAGCCAATGCTCATGCTCAT300                         GATTTCTCAGAAAGCCTCACAAAAGGTTTGCACGATTCAACTGGTGTACA350                         TTTGATTCATGCCACAGAACCGACATTGTCACGAAGTCCGGAAGTGGAAG400                         GTACGGATGGCGATTTGGATGCCTTAATTCGTAAAGCACAACATGAACCA450                         ATTACTACAGCGATGGCCGATCCTTCCTTAGATGCATCGCTTCCTGACAA500                         TGTTACGATAATGAGAACTACCATGCAACCTAGTTTTTTAATTTCGTTTA550                         TGGTGGATGCACGTGGAGGAGCAATGCGTGGTTGTAGGCATTCCGGTGTC600                         AGAATCATTATACCACCGAGGAAAGCGCCGCAACCTACACGGGTCACATG650                         CAGATACCTTGGAAAGGACAAGTTAGCGCATCCACCACCATTAAGTGAAG700                         GTGAAGCGCTCGCNTCACGTATACTTGAAATGGCACCACATGGAGCAAAA750                         TTCTTAGGCCCTGTTATATTGGAAGTACCACATTTTGCATCACTTCGTGG800                         ACGAGAGAGAGAGATTGTCATTTTGCGTTCTGATGATGGGCAGCATTGGA850                         AAGAGCATCAGCTTGAAGCAACAGAAGATGCTGTACAAGAGGTGCTCAAT900                         GAATCGTTTGATGCAGAAGAGTTGTCGCAACTTGATGATTTGCATACATC950                         ACGGATTACGCGTATCCTGACCAATGATTTCCCAATGTATTTCGCGGTCG1000                        TTACTCGTGTGCGGCAAGAAGTGCACTGTGTTGGTCCAGAAGGTGGTGTA1050                        ATACTCTCTTCAGTTGTTCCTCATGTGCAGGCCATATTTCCGGATGGTTC1100                        CTTGACTAAGACGATCAAAGTATCTGTGCAAGCCCAGCCAGTTCCACAAG1150                        AGATAGTCACTCGTTTACATGGGAATAGAGTCGCTGTTTCTCCAATTGTA1200                        ACTGTTGAACCGCGTCGTCGCAAATTC1227                                               (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1227 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      GAATTTGCGACGACGCGGTTCAACAGTTACAATTGGAGAAACAGCGACTC50                          TATTCCCATGTAAACGAGTGACTATCTCTTGTGGAACTGGCTGGGCTTGC100                         ACAGATACTTTGATCGTCTTAGTCAAGGAACCATCCGGAAATATGGCCTG150                         CACATGAGGAACAACTGAAGAGAGTATTACACCACCTTCTGGACCAACAC200                         AGTGCACTTCTTGCCGCACACGAGTAACGACCGCGAAATACATTGGGAAA250                         TCATTGGTCAGGATACGCGTAATCCGTGATGTATGCAAATCATCAAGTTG300                         CGACAACTCTTCTGCATCAAACGATTCATTGAGCACCTCTTGTACAGCAT350                         CTTCTGTTGCTTCAAGCTGATGCTCTTTCCAATGCTGCCCATCATCAGAA400                         CGCAAAATGACAATCTCTCTCTCTCGTCCACGAAGTGATGCAAAATGTGG450                         TACTTCCAATATAACAGGGCCTAAGAATTTTGCTCCATGTGGTGCCATTT500                         CAAGTATACGTGANGCGAGCGCTTCACCTTCACTTAATGGTGGTGGATGC550                         GCTAACTTGTCCTTTCCAAGGTATCTGCATGTGACCCGTGTAGGTTGCGG600                         CGCTTTCCTCGGTGGTATAATGATTCTGACACCGGAATGCCTACAACCAC650                         GCATTGCTCCTCCACGTGCATCCACCATAAACGAAATTAAAAAACTAGGT700                         TGCATGGTAGTTCTCATTATCGTAACATTGTCAGGAAGCGATGCATCTAA750                         GGAAGGATCGGCCATCGCTGTAGTAATTGGTTCATGTTGTGCTTTACGAA800                         TTAAGGCATCCAAATCGCCATCCGTACCTTCCACTTCCGGACTTCGTGAC850                         AATGTCGGTTCTGTGGCATGAATCAAATGTACACCAGTTGAATCGTGCAA900                         ACCTTTTGTGAGGCTTTCTGAGAAATCATGAGCATGAGCATTGGCTGTGA950                         TCTGATTATCTTCACCTTCATCTTCGGAATCGGAAAACATGGTTTCATTC1000                        ATTGCTTCGGGATTCTGAGGTTTATATCTTTCTTCAGTAACGGTTGTGGT1050                        CTCCGTTATCACAGTAGTCTCGGTAATTGTTTTAAGCGCTTCAACCACGG1100                        ATACATACCCTAGACGTTCAGCAATCGATAATGGAGTTTGTCCTGTCGAT1150                        GTATGAACATTTGGACTTGCACCATGTTCCAACAAGTAACGTACAACACT1200                        GTTATGCCCTTGCTGAGCAGCTTGATG1227                                               (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 573 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..573                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      AATAACGACAAGGTAGCATTGTTACTTCTAGAAAATGGTGCT42                                  AsnAsnAspLysValAlaLeuLeuLeuLeuGluAsnGlyAla                                    1510                                                                          TCTGCACATGCCGCTGCCAAGAATGGGTACACTCCTTTACAT84                                  SerAlaHisAlaAlaAlaLysAsnGlyTyrThrProLeuHis                                    152025                                                                        ATTGCCGCGAAGAAGAATCAGATGGATATTGCTAGCACTCTC126                                 IleAlaAlaLysLysAsnGlnMetAspIleAlaSerThrLeu                                    303540                                                                        CTTCATTATAAGGCAAATGCGAATGCTGAAAGCAAAGCTGGC168                                 LeuHisTyrLysAlaAsnAlaAsnAlaGluSerLysAlaGly                                    455055                                                                        TTTACACCACTTCATCTTGCCGCCCAGGAGGGCCATCGCGAA210                                 PheThrProLeuHisLeuAlaAlaGlnGluGlyHisArgGlu                                    606570                                                                        ATGGCTGCGTTATTAATTGAAAATGGAGCAAAAGTTGGAGCT252                                 MetAlaAlaLeuLeuIleGluAsnGlyAlaLysValGlyAla                                    7580                                                                          CAGGCAAGGAATGGCTTGACACCAATGCATTTATGTGCACAG294                                 GlnAlaArgAsnGlyLeuThrProMetHisLeuCysAlaGln                                    859095                                                                        GAGGATCGTGTGAGCGTAGCAGAAGAACTAGTGAAAGAAAAC336                                 GluAspArgValSerValAlaGluGluLeuValLysGluAsn                                    100105110                                                                     GCAGCCATTGATCCCAAAACGAAAGCAGGATATACGCCGTTA378                                 AlaAlaIleAspProLysThrLysAlaGlyTyrThrProLeu                                    115120125                                                                     CATGTTGCTTGCCATTTTGGACAAATAAACATGGTCCGTTTC420                                 HisValAlaCysHisPheGlyGlnIleAsnMetValArgPhe                                    130135140                                                                     TTGATTGAGCATGGCGCACGAGTTTCAGTTATTACTCGTGCT462                                 LeuIleGluHisGlyAlaArgValSerValIleThrArgAla                                    145150                                                                        TCCTATACTCCTCTGCATCAAGCTGCTCAGCAAGGGCATAAC504                                 SerTyrThrProLeuHisGlnAlaAlaGlnGlnGlyHisAsn                                    155160165                                                                     AGTGTTGTACGTTACTTGTTGGAACATGGTGCAAGTCCAAAT546                                 SerValValArgTyrLeuLeuGluHisGlyAlaSerProAsn                                    170175180                                                                     GTTCATACATCGACAGGACAAACTCCA573                                                ValHisThrSerThrGlyGlnThrPro                                                   185190                                                                        (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 191 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      AsnAsnAspLysValAlaLeuLeuLeuLeuGluAsnGlyAla                                    1510                                                                          SerAlaHisAlaAlaAlaLysAsnGlyTyrThrProLeuHis                                    152025                                                                        IleAlaAlaLysLysAsnGlnMetAspIleAlaSerThrLeu                                    303540                                                                        LeuHisTyrLysAlaAsnAlaAsnAlaGluSerLysAlaGly                                    455055                                                                        PheThrProLeuHisLeuAlaAlaGlnGluGlyHisArgGlu                                    606570                                                                        MetAlaAlaLeuLeuIleGluAsnGlyAlaLysValGlyAla                                    7580                                                                          GlnAlaArgAsnGlyLeuThrProMetHisLeuCysAlaGln                                    859095                                                                        GluAspArgValSerValAlaGluGluLeuValLysGluAsn                                    100105110                                                                     AlaAlaIleAspProLysThrLysAlaGlyTyrThrProLeu                                    115120125                                                                     HisValAlaCysHisPheGlyGlnIleAsnMetValArgPhe                                    130135140                                                                     LeuIleGluHisGlyAlaArgValSerValIleThrArgAla                                    145150                                                                        SerTyrThrProLeuHisGlnAlaAlaGlnGlnGlyHisAsn                                    155160165                                                                     SerValValArgTyrLeuLeuGluHisGlyAlaSerProAsn                                    170175180                                                                     ValHisThrSerThrGlyGlnThrPro                                                   185190                                                                        (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 573 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      TGGAGTTTGTCCTGTCGATGTATGAACATTTGGACTTGCACCATGTTCCA50                          ACAAGTAACGTACAACACTGTTATGCCCTTGCTGAGCAGCTTGATGCAGA100                         GGAGTATAGGAAGCACGAGTAATAACTGAAACTCGTGCGCCATGCTCAAT150                         CAAGAAACGGACCATGTTTATTTGTCCAAAATGGCAAGCAACATGTAACG200                         GCGTATATCCTGCTTTCGTTTTGGGATCAATGGCTGCGTTTTCTTTCACT250                         AGTTCTTCTGCTACGCTCACACGATCCTCCTGTGCACATAAATGCATTGG300                         TGTCAAGCCATTCCTTGCCTGAGCTCCAACTTTTGCTCCATTTTCAATTA350                         ATAACGCAGCCATTTCGCGATGGCCCTCCTGGGCGGCAAGATGAAGTGGT400                         GTAAAGCCAGCTTTGCTTTCAGCATTCGCATTTGCCTTATAATGAAGGAG450                         AGTGCTAGCAATATCCATCTGATTCTTCTTCGCGGCAATATGTAAAGGAG500                         TGTACCCATTCTTGGCAGCGGCATGTGCAGAAGCACCATTTTCTAGAAGT550                         AACAATGCTACCTTGTCGTTATT573                                                    (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 911 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..909                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      GTGGATGATGTTACTGTTGACTATCTCACTCCTCTTCATGTG42                                  ValAspAspValThrValAspTyrLeuThrProLeuHisVal                                    1510                                                                          GCTGCTCATTGCGGACATGTCCGTGTCGCTAAACTTTTGCTG84                                  AlaAlaHisCysGlyHisValArgValAlaLysLeuLeuLeu                                    152025                                                                        GATCGTAATGCTGACCCGAATGCTCGAGCTCTCAATGGCTTC126                                 AspArgAsnAlaAspProAsnAlaArgAlaLeuAsnGlyPhe                                    303540                                                                        ACACCGCTGCATATCGCTTGCAAAAAAAATCGCATTAAAATT168                                 ThrProLeuHisIleAlaCysLysLysAsnArgIleLysIle                                    455055                                                                        GTCGAACTGCTACTGAAATACCACGCTGCAATCGAAGCAACT210                                 ValGluLeuLeuLeuLysTyrHisAlaAlaIleGluAlaThr                                    606570                                                                        ACTGAATCCGGTCTCTCACCGCTGCATGTCGCTGCTTTTATG252                                 ThrGluSerGlyLeuSerProLeuHisValAlaAlaPheMet                                    7580                                                                          GGTGCTATAAACATTGTCATCTATTTACTACAACAAGGTGCT294                                 GlyAlaIleAsnIleValIleTyrLeuLeuGlnGlnGlyAla                                    859095                                                                        AATGCAGATGTGGCTACAGTACGCGGTGAAACGCCTCTTCAT336                                 AsnAlaAspValAlaThrValArgGlyGluThrProLeuHis                                    100105110                                                                     TTAGCTGCACGAGCAAACCAAACGGACATTGTTCGTGTTTTG378                                 LeuAlaAlaArgAlaAsnGlnThrAspIleValArgValLeu                                    115120125                                                                     GTGCGTAATGGAGCACAGGTGGATGCTGCTGCTCGTGAACTA420                                 ValArgAsnGlyAlaGlnValAspAlaAlaAlaArgGluLeu                                    130135140                                                                     CAAACTCCACTGCACATTGCATCACGTCTTGGTAATACCGAC462                                 GlnThrProLeuHisIleAlaSerArgLeuGlyAsnThrAsp                                    145150                                                                        ATCGTCATTTTGTTGCTGCAGGCTAATGCATCACCAAATGCT504                                 IleValIleLeuLeuLeuGlnAlaAsnAlaSerProAsnAla                                    155160165                                                                     GCCACAAGAGATCTTTATACTCCTCTTCATATTGCTGCCAAG546                                 AlaThrArgAspLeuTyrThrProLeuHisIleAlaAlaLys                                    170175180                                                                     GAGGGGCAAGAGGAAGTGGCAGCAATATTGATGGATCATGGA588                                 GluGlyGlnGluGluValAlaAlaIleLeuMetAspHisGly                                    185190195                                                                     ACCGACAAGACACTGCTCACGAAAAAGGGTTTTACGCCGTTG630                                 ThrAspLysThrLeuLeuThrLysLysGlyPheThrProLeu                                    200205210                                                                     CATTTAGCTGCTAAGTATGGCAATTTGCCGGTCGCGAAATCA672                                 HisLeuAlaAlaLysTyrGlyAsnLeuProValAlaLysSer                                    215220                                                                        TTGCTAGAACGAGGAACACCGGTTGACATTGAAGGCAAGAAT714                                 LeuLeuGluArgGlyThrProValAspIleGluGlyLysAsn                                    225230235                                                                     CAGGTAACACCTCTGCATGTAGCGGCACATTACAATAACGAC756                                 GlnValThrProLeuHisValAlaAlaHisTyrAsnAsnAsp                                    240245250                                                                     AAGGTAGCATTGTTACTTCTAGAAAATGGTGCTTCTGCACAT798                                 LysValAlaLeuLeuLeuLeuGluAsnGlyAlaSerAlaHis                                    255260265                                                                     GCCGCTGCCAAGAATGGGTACACTCCTTTACATATTGCCGCG840                                 AlaAlaAlaLysAsnGlyTyrThrProLeuHisIleAlaAla                                    270275280                                                                     AAGAAGAATCAGATGGATATTGCTAGCACTCTCCTTCATTAT882                                 LysLysAsnGlnMetAspIleAlaSerThrLeuLeuHisTyr                                    285290                                                                        AAGGCAAATGCGAATGCTGAAAGCAAAGC911                                              LysAlaAsnAlaAsnAlaGluSerLys                                                   295300                                                                        (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 303 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                      ValAspAspValThrValAspTyrLeuThrProLeuHisVal                                    1510                                                                          AlaAlaHisCysGlyHisValArgValAlaLysLeuLeuLeu                                    152025                                                                        AspArgAsnAlaAspProAsnAlaArgAlaLeuAsnGlyPhe                                    303540                                                                        ThrProLeuHisIleAlaCysLysLysAsnArgIleLysIle                                    455055                                                                        ValGluLeuLeuLeuLysTyrHisAlaAlaIleGluAlaThr                                    606570                                                                        ThrGluSerGlyLeuSerProLeuHisValAlaAlaPheMet                                    7580                                                                          GlyAlaIleAsnIleValIleTyrLeuLeuGlnGlnGlyAla                                    859095                                                                        AsnAlaAspValAlaThrValArgGlyGluThrProLeuHis                                    100105110                                                                     LeuAlaAlaArgAlaAsnGlnThrAspIleValArgValLeu                                    115120125                                                                     ValArgAsnGlyAlaGlnValAspAlaAlaAlaArgGluLeu                                    130135140                                                                     GlnThrProLeuHisIleAlaSerArgLeuGlyAsnThrAsp                                    145150                                                                        IleValIleLeuLeuLeuGlnAlaAsnAlaSerProAsnAla                                    155160165                                                                     AlaThrArgAspLeuTyrThrProLeuHisIleAlaAlaLys                                    170175180                                                                     GluGlyGlnGluGluValAlaAlaIleLeuMetAspHisGly                                    185190195                                                                     ThrAspLysThrLeuLeuThrLysLysGlyPheThrProLeu                                    200205210                                                                     HisLeuAlaAlaLysTyrGlyAsnLeuProValAlaLysSer                                    215220                                                                        LeuLeuGluArgGlyThrProValAspIleGluGlyLysAsn                                    225230235                                                                     GlnValThrProLeuHisValAlaAlaHisTyrAsnAsnAsp                                    240245250                                                                     LysValAlaLeuLeuLeuLeuGluAsnGlyAlaSerAlaHis                                    255260265                                                                     AlaAlaAlaLysAsnGlyTyrThrProLeuHisIleAlaAla                                    270275280                                                                     LysLysAsnGlnMetAspIleAlaSerThrLeuLeuHisTyr                                    285290                                                                        LysAlaAsnAlaAsnAlaGluSerLys                                                   295300                                                                        (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 911 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      GCTTTGCTTTCAGCATTCGCATTTGCCTTATAATGAAGGAGAGTGCTAGC50                          AATATCCATCTGATTCTTCTTCGCGGCAATATGTAAAGGAGTGTACCCAT100                         TCTTGGCAGCGGCATGTGCAGAAGCACCATTTTCTAGAAGTAACAATGCT150                         ACCTTGTCGTTATTGTAATGTGCCGCTACATGCAGAGGTGTTACCTGATT200                         CTTGCCTTCAATGTCAACCGGTGTTCCTCGTTCTAGCAATGATTTCGCGA250                         CCGGCAAATTGCCATACTTAGCAGCTAAATGCAACGGCGTAAAACCCTTT300                         TTCGTGAGCAGTGTCTTGTCGGTTCCATGATCCATCAATATTGCTGCCAC350                         TTCCTCTTGCCCCTCCTTGGCAGCAATATGAAGAGGAGTATAAAGATCTC400                         TTGTGGCAGCATTTGGTGATGCATTAGCCTGCAGCAACAAAATGACGATG450                         TCGGTATTACCAAGACGTGATGCAATGTGCAGTGGAGTTTGTAGTTCACG500                         AGCAGCAGCATCCACCTGTGCTCCATTACGCACCAAAACACGAACAATGT550                         CCGTTTGGTTTGCTCGTGCAGCTAAATGAAGAGGCGTTTCACCGCGTACT600                         GTAGCCACATCTGCATTAGCACCTTGTTGTAGTAAATAGATGACAATGTT650                         TATAGCACCCATAAAAGCAGCGACATGCAGCGGTGAGAGACCGGATTCAG700                         TAGTTGCTTCGATTGCAGCGTGGTATTTCAGTAGCAGTTCGACAATTTTA750                         ATGCGATTTTTTTTGCAAGCGATATGCAGCGGTGTGAAGCCATTGAGAGC800                         TCGAGCATTCGGGTCAGCATTACGATCCAGCAAAAGTTTAGCGACACGGA850                         CATGTCCGCAATGAGCAGCCACATGAAGAGGAGTGAGATAGTCAACAGTA900                         ACATCATCCAC911                                                                (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 909 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      GTGGATGATGTTACTGTTGACTATCTCACTCCTCTTCATGTGGCTGCTCA50                          TTGCGGACATGTCCGTGTCGCTAAACTTTTGCTGGATCGTAATGCTGACC100                         CGAATGCTCGAGCTCTCAATGGCTTCACACCGCTGCATATCGCTTGCAAA150                         AAAAATCGCATTAAAATTGTCGAACTGCTACTGAAATACCACGCTGCAAT200                         CGAAGCAACTACTGAATCCGGTCTCTCACCGCTGCATGTCGCTGCTTTTA250                         TGGGTGCTATAAACATTGTCATCTATTTACTACAACAAGGTGCTAATGCA300                         GATGTGGCTACAGTACGCGGTGAAACGCCTCTTCATTTAGCTGCACGAGC350                         AAACCAAACGGACATTGTTCGTGTTTTGGTGCGTAATGGAGCACAGGTGG400                         ATGCTGCTGCTCGTGAACTACAAACTCCACTGCACATTGCATCACGTCTT450                         GGTAATACCGACATCGTCATTTTGTTGCTGCAGGCTAATGCATCACCAAA500                         TGCTGCCACAAGAGATCTTTATACTCCTCTTCATATTGCTGCCAAGGAGG550                         GGCAAGAGGAAGTGGCAGCAATATTGATGGATCATGGAACCGACAAGACA600                         CTGCTCACGAAAAAGGGTTTTACGCCGTTGCATTTAGCTGCTAAGTATGG650                         CAATTTGCCGGTCGCGAAATCATTGCTAGAACGAGGAACACCGGTTGACA700                         TTGAAGGCAAGAATCAGGTAACACCTCTGCATGTAGCGGCACATTACAAT750                         AACGACAAGGTAGCATTGTTACTTCTAGAAAATGGTGCTTCTGCACATGC800                         CGCTGCCAAGAATGGGTACACTCCTTTACATATTGCCGCGAAGAAGAATC850                         AGATGGATATTGCTAGCACTCTCCTTCATTATAAGGCAAATGCGAATGCT900                         GAAAGCAAA909                                                                  (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 909 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      TTTGCTTTCAGCATTCGCATTTGCCTTATAATGAAGGAGAGTGCTAGCAA50                          TATCCATCTGATTCTTCTTCGCGGCAATATGTAAAGGAGTGTACCCATTC100                         TTGGCAGCGGCATGTGCAGAAGCACCATTTTCTAGAAGTAACAATGCTAC150                         CTTGTCGTTATTGTAATGTGCCGCTACATGCAGAGGTGTTACCTGATTCT200                         TGCCTTCAATGTCAACCGGTGTTCCTCGTTCTAGCAATGATTTCGCGACC250                         GGCAAATTGCCATACTTAGCAGCTAAATGCAACGGCGTAAAACCCTTTTT300                         CGTGAGCAGTGTCTTGTCGGTTCCATGATCCATCAATATTGCTGCCACTT350                         CCTCTTGCCCCTCCTTGGCAGCAATATGAAGAGGAGTATAAAGATCTCTT400                         GTGGCAGCATTTGGTGATGCATTAGCCTGCAGCAACAAAATGACGATGTC450                         GGTATTACCAAGACGTGATGCAATGTGCAGTGGAGTTTGTAGTTCACGAG500                         CAGCAGCATCCACCTGTGCTCCATTACGCACCAAAACACGAACAATGTCC550                         GTTTGGTTTGCTCGTGCAGCTAAATGAAGAGGCGTTTCACCGCGTACTGT600                         AGCCACATCTGCATTAGCACCTTGTTGTAGTAAATAGATGACAATGTTTA650                         TAGCACCCATAAAAGCAGCGACATGCAGCGGTGAGAGACCGGATTCAGTA700                         GTTGCTTCGATTGCAGCGTGGTATTTCAGTAGCAGTTCGACAATTTTAAT750                         GCGATTTTTTTTGCAAGCGATATGCAGCGGTGTGAAGCCATTGAGAGCTC800                         GAGCATTCGGGTCAGCATTACGATCCAGCAAAAGTTTAGCGACACGGACA850                         TGTCCGCAATGAGCAGCCACATGAAGAGGAGTGAGATAGTCAACAGTAAC900                         ATCATCCAC909                                                                  (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1096 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 51..1094                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      GGTTTAATTACCCAAGTTTGAGGCGGCTGACTGATATAACTCAACTATTG50                          ATGAGTAATCCTATAGTCGAGGGAAGTGGCTGGCCCGCAGAA92                                  MetSerAsnProIleValGluGlySerGlyTrpProAlaGlu                                    1510                                                                          CCAAAAGATTCACAACATCAACAACAAATTCCTGATGATAAC134                                 ProLysAspSerGlnHisGlnGlnGlnIleProAspAspAsn                                    152025                                                                        AGTCAACATTCCAACAAAGGTGAGAGCAGTGCAAGTTTTTTA176                                 SerGlnHisSerAsnLysGlyGluSerSerAlaSerPheLeu                                    303540                                                                        CGAGCAGCAAGAGCTGGAAATTTGGATCGTGTACTTGAACTA218                                 ArgAlaAlaArgAlaGlyAsnLeuAspArgValLeuGluLeu                                    455055                                                                        CTTCGTTCGGGCACCGATATCAACACATGCAATGCGAATGGC260                                 LeuArgSerGlyThrAspIleAsnThrCysAsnAlaAsnGly                                    606570                                                                        CTTAATGCATTGCATCTGGCCTCCAAAGAAGGTCATCATGAA302                                 LeuAsnAlaLeuHisLeuAlaSerLysGluGlyHisHisGlu                                    7580                                                                          GTGGTCCGCGAACTTCTGAAAAGAAAAGCAGATGTTGATGCT344                                 ValValArgGluLeuLeuLysArgLysAlaAspValAspAla                                    859095                                                                        GCCACTAGAAAGGGTAACACAGCGTTACATATAGCATCATTG386                                 AlaThrArgLysGlyAsnThrAlaLeuHisIleAlaSerLeu                                    100105110                                                                     GCAGGACAAGAACTAATCGTCACAGTACTTGTTGAAAATGGT428                                 AlaGlyGlnGluLeuIleValThrValLeuValGluAsnGly                                    115120125                                                                     GCTAATGTTAACGTACAATCACTAAACGGTTTTACACCACTT470                                 AlaAsnValAsnValGlnSerLeuAsnGlyPheThrProLeu                                    130135140                                                                     TACATGGCTGCACAAGAAAATCACGAATCTGTTGTACGCTAT512                                 TyrMetAlaAlaGlnGluAsnHisGluSerValValArgTyr                                    145150                                                                        CTTCTTGCCCACAATGCCAATCAAGCTTTAAGTACAGAAGAC554                                 LeuLeuAlaHisAsnAlaAsnGlnAlaLeuSerThrGluAsp                                    155160165                                                                     GGTTTTACGCCACTGGCAGTTGCCTTGCAACAAGGTCACGAT596                                 GlyPheThrProLeuAlaValAlaLeuGlnGlnGlyHisAsp                                    170175180                                                                     CGTGTGGTCGCTGTTTTGCTTGAAAATGACACGCGCGGGAAA638                                 ArgValValAlaValLeuLeuGluAsnAspThrArgGlyLys                                    185190195                                                                     GTGCGCTTGCCAGCACTGCATATTGCTGCTAAAAAAGATGAT680                                 ValArgLeuProAlaLeuHisIleAlaAlaLysLysAspAsp                                    200205210                                                                     ACGAAAGCAGCTACGCTATTACTTCAAAATGAGCATAACTCG722                                 ThrLysAlaAlaThrLeuLeuLeuGlnAsnGluHisAsnSer                                    215220                                                                        GATGTGACTTCGAAAAGCGGCTTTACTCCGCTTCATATCGCC764                                 AspValThrSerLysSerGlyPheThrProLeuHisIleAla                                    225230235                                                                     GCTCACTATGGAAATGAGAACGTAGCACAACTGCTACTCGAA806                                 AlaHisTyrGlyAsnGluAsnValAlaGlnLeuLeuLeuGlu                                    240245250                                                                     AAGGGAGCCAATGTGAATTACCAAGCGAGACATAACATAAGT848                                 LysGlyAlaAsnValAsnTyrGlnAlaArgHisAsnIleSer                                    255260265                                                                     CCGTTACACGTTGCAACAAAATGGGGTCGTACAAACATGGTT890                                 ProLeuHisValAlaThrLysTrpGlyArgThrAsnMetVal                                    270275280                                                                     TCGTTATTGTTGGCTCATGGGGCCGTAATTGACTGTCGCACA932                                 SerLeuLeuLeuAlaHisGlyAlaValIleAspCysArgThr                                    285290                                                                        CGTGATTTACTAACACCATTACACTGTGCTTCTCGTTCAGGT974                                 ArgAspLeuLeuThrProLeuHisCysAlaSerArgSerGly                                    295300305                                                                     CATGATCAAGTTGTTGATTTGTTGCTTGAAAAAGGAGCTCCA1016                                HisAspGlnValValAspLeuLeuLeuGluLysGlyAlaPro                                    310315320                                                                     ATCAGTGCTAAGACAAAAAATGGTTTGGCTCCCTTACATATG1058                                IleSerAlaLysThrLysAsnGlyLeuAlaProLeuHisMet                                    325330335                                                                     GCAGCACAGGTGGATGATGTTACTGTTGACTATCTCAC1096                                    AlaAlaGlnValAspAspValThrValAspTyrLeu                                          340345                                                                        (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 348 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      MetSerAsnProIleValGluGlySerGlyTrpProAlaGlu                                    1510                                                                          ProLysAspSerGlnHisGlnGlnGlnIleProAspAspAsn                                    152025                                                                        SerGlnHisSerAsnLysGlyGluSerSerAlaSerPheLeu                                    303540                                                                        ArgAlaAlaArgAlaGlyAsnLeuAspArgValLeuGluLeu                                    455055                                                                        LeuArgSerGlyThrAspIleAsnThrCysAsnAlaAsnGly                                    606570                                                                        LeuAsnAlaLeuHisLeuAlaSerLysGluGlyHisHisGlu                                    7580                                                                          ValValArgGluLeuLeuLysArgLysAlaAspValAspAla                                    859095                                                                        AlaThrArgLysGlyAsnThrAlaLeuHisIleAlaSerLeu                                    100105110                                                                     AlaGlyGlnGluLeuIleValThrValLeuValGluAsnGly                                    115120125                                                                     AlaAsnValAsnValGlnSerLeuAsnGlyPheThrProLeu                                    130135140                                                                     TyrMetAlaAlaGlnGluAsnHisGluSerValValArgTyr                                    145150                                                                        LeuLeuAlaHisAsnAlaAsnGlnAlaLeuSerThrGluAsp                                    155160165                                                                     GlyPheThrProLeuAlaValAlaLeuGlnGlnGlyHisAsp                                    170175180                                                                     ArgValValAlaValLeuLeuGluAsnAspThrArgGlyLys                                    185190195                                                                     ValArgLeuProAlaLeuHisIleAlaAlaLysLysAspAsp                                    200205210                                                                     ThrLysAlaAlaThrLeuLeuLeuGlnAsnGluHisAsnSer                                    215220                                                                        AspValThrSerLysSerGlyPheThrProLeuHisIleAla                                    225230235                                                                     AlaHisTyrGlyAsnGluAsnValAlaGlnLeuLeuLeuGlu                                    240245250                                                                     LysGlyAlaAsnValAsnTyrGlnAlaArgHisAsnIleSer                                    255260265                                                                     ProLeuHisValAlaThrLysTrpGlyArgThrAsnMetVal                                    270275280                                                                     SerLeuLeuLeuAlaHisGlyAlaValIleAspCysArgThr                                    285290                                                                        ArgAspLeuLeuThrProLeuHisCysAlaSerArgSerGly                                    295300305                                                                     HisAspGlnValValAspLeuLeuLeuGluLysGlyAlaPro                                    310315320                                                                     IleSerAlaLysThrLysAsnGlyLeuAlaProLeuHisMet                                    325330335                                                                     AlaAlaGlnValAspAspValThrValAspTyrLeu                                          340345                                                                        (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1096 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      GTGAGATAGTCAACAGTAACATCATCCACCTGTGCTGCCATATGTAAGGG50                          AGCCAAACCATTTTTTGTCTTAGCACTGATTGGAGCTCCTTTTTCAAGCA100                         ACAAATCAACAACTTGATCATGACCTGAACGAGAAGCACAGTGTAATGGT150                         GTTAGTAAATCACGTGTGCGACAGTCAATTACGGCCCCATGAGCCAACAA200                         TAACGAAACCATGTTTGTACGACCCCATTTTGTTGCAACGTGTAACGGAC250                         TTATGTTATGTCTCGCTTGGTAATTCACATTGGCTCCCTTTTCGAGTAGC300                         AGTTGTGCTACGTTCTCATTTCCATAGTGAGCGGCGATATGAAGCGGAGT350                         AAAGCCGCTTTTCGAAGTCACATCCGAGTTATGCTCATTTTGAAGTAATA400                         GCGTAGCTGCTTTCGTATCATCTTTTTTAGCAGCAATATGCAGTGCTGGC450                         AAGCGCACTTTCCCGCGCGTGTCATTTTCAAGCAAAACAGCGACCACACG500                         ATCGTGACCTTGTTGCAAGGCAACTGCCAGTGGCGTAAAACCGTCTTCTG550                         TACTTAAAGCTTGATTGGCATTGTGGGCAAGAAGATAGCGTACAACAGAT600                         TCGTGATTTTCTTGTGCAGCCATGTAAAGTGGTGTAAAACCGTTTAGTGA650                         TTGTACGTTAACATTAGCACCATTTTCAACAAGTACTGTGACGATTAGTT700                         CTTGTCCTGCCAATGATGCTATATGTAACGCTGTGTTACCCTTTCTAGTG750                         GCAGCATCAACATCTGCTTTTCTTTTCAGAAGTTCGCGGACCACTTCATG800                         ATGACCTTCTTTGGAGGCCAGATGCAATGCATTAAGGCCATTCGCATTGC850                         ATGTGTTGATATCGGTGCCCGAACGAAGTAGTTCAAGTACACGATCCAAA900                         TTTCCAGCTCTTGCTGCTCGTAAAAAACTTGCACTGCTCTCACCTTTGTT950                         GGAATGTTGACTGTTATCATCAGGAATTTGTTGTTGATGTTGTGAATCTT1000                        TTGGTTCTGCGGGCCAGCCACTTCCCTCGACTATAGGATTACTCATCAAT1050                        AGTTGAGTTATATCAGTCAGCCGCCTCAAACTTGGGTAATTAAACC1096                            (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1044 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      ATGAGTAATCCTATAGTCGAGGGAAGTGGCTGGCCCGCAGAACCAAAAGA50                          TTCACAACATCAACAACAAATTCCTGATGATAACAGTCAACATTCCAACA100                         AAGGTGAGAGCAGTGCAAGTTTTTTACGAGCAGCAAGAGCTGGAAATTTG150                         GATCGTGTACTTGAACTACTTCGTTCGGGCACCGATATCAACACATGCAA200                         TGCGAATGGCCTTAATGCATTGCATCTGGCCTCCAAAGAAGGTCATCATG250                         AAGTGGTCCGCGAACTTCTGAAAAGAAAAGCAGATGTTGATGCTGCCACT300                         AGAAAGGGTAACACAGCGTTACATATAGCATCATTGGCAGGACAAGAACT350                         AATCGTCACAGTACTTGTTGAAAATGGTGCTAATGTTAACGTACAATCAC400                         TAAACGGTTTTACACCACTTTACATGGCTGCACAAGAAAATCACGAATCT450                         GTTGTACGCTATCTTCTTGCCCACAATGCCAATCAAGCTTTAAGTACAGA500                         AGACGGTTTTACGCCACTGGCAGTTGCCTTGCAACAAGGTCACGATCGTG550                         TGGTCGCTGTTTTGCTTGAAAATGACACGCGCGGGAAAGTGCGCTTGCCA600                         GCACTGCATATTGCTGCTAAAAAAGATGATACGAAAGCAGCTACGCTATT650                         ACTTCAAAATGAGCATAACTCGGATGTGACTTCGAAAAGCGGCTTTACTC700                         CGCTTCATATCGCCGCTCACTATGGAAATGAGAACGTAGCACAACTGCTA750                         CTCGAAAAGGGAGCCAATGTGAATTACCAAGCGAGACATAACATAAGTCC800                         GTTACACGTTGCAACAAAATGGGGTCGTACAAACATGGTTTCGTTATTGT850                         TGGCTCATGGGGCCGTAATTGACTGTCGCACACGTGATTTACTAACACCA900                         TTACACTGTGCTTCTCGTTCAGGTCATGATCAAGTTGTTGATTTGTTGCT950                         TGAAAAAGGAGCTCCAATCAGTGCTAAGACAAAAAATGGTTTGGCTCCCT1000                        TACATATGGCAGCACAGGTGGATGATGTTACTGTTGACTATCTC1044                              (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1044 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      GAGATAGTCAACAGTAACATCATCCACCTGTGCTGCCATATGTAAGGGAG50                          CCAAACCATTTTTTGTCTTAGCACTGATTGGAGCTCCTTTTTCAAGCAAC100                         AAATCAACAACTTGATCATGACCTGAACGAGAAGCACAGTGTAATGGTGT150                         TAGTAAATCACGTGTGCGACAGTCAATTACGGCCCCATGAGCCAACAATA200                         ACGAAACCATGTTTGTACGACCCCATTTTGTTGCAACGTGTAACGGACTT250                         ATGTTATGTCTCGCTTGGTAATTCACATTGGCTCCCTTTTCGAGTAGCAG300                         TTGTGCTACGTTCTCATTTCCATAGTGAGCGGCGATATGAAGCGGAGTAA350                         AGCCGCTTTTCGAAGTCACATCCGAGTTATGCTCATTTTGAAGTAATAGC400                         GTAGCTGCTTTCGTATCATCTTTTTTAGCAGCAATATGCAGTGCTGGCAA450                         GCGCACTTTCCCGCGCGTGTCATTTTCAAGCAAAACAGCGACCACACGAT500                         CGTGACCTTGTTGCAAGGCAACTGCCAGTGGCGTAAAACCGTCTTCTGTA550                         CTTAAAGCTTGATTGGCATTGTGGGCAAGAAGATAGCGTACAACAGATTC600                         GTGATTTTCTTGTGCAGCCATGTAAAGTGGTGTAAAACCGTTTAGTGATT650                         GTACGTTAACATTAGCACCATTTTCAACAAGTACTGTGACGATTAGTTCT700                         TGTCCTGCCAATGATGCTATATGTAACGCTGTGTTACCCTTTCTAGTGGC750                         AGCATCAACATCTGCTTTTCTTTTCAGAAGTTCGCGGACCACTTCATGAT800                         GACCTTCTTTGGAGGCCAGATGCAATGCATTAAGGCCATTCGCATTGCAT850                         GTGTTGATATCGGTGCCCGAACGAAGTAGTTCAAGTACACGATCCAAATT900                         TCCAGCTCTTGCTGCTCGTAAAAAACTTGCACTGCTCTCACCTTTGTTGG950                         AATGTTGACTGTTATCATCAGGAATTTGTTGTTGATGTTGTGAATCTTTT1000                        GGTTCTGCGGGCCAGCCACTTCCCTCGACTATAGGATTACTCAT1044                              (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5503 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 51..5285                                                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      GGTTTAATTACCCAAGTTTGAGGCGGCTGACTGATATAACTCAACTATTG50                          ATGAGTAATCCTATAGTCGAGGGAAGTGGCTGGCCCGCAGAA92                                  MetSerAsnProIleValGluGlySerGlyTrpProAlaGlu                                    1510                                                                          CCAAAAGATTCACAACATCAACAACAAATTCCTGATGATAAC134                                 ProLysAspSerGlnHisGlnGlnGlnIleProAspAspAsn                                    152025                                                                        AGTCAACATTCCAACAAAGGTGAGAGCAGTGCAAGTTTTTTA176                                 SerGlnHisSerAsnLysGlyGluSerSerAlaSerPheLeu                                    303540                                                                        CGAGCAGCAAGAGCTGGAAATTTGGATCGTGTACTTGAACTA218                                 ArgAlaAlaArgAlaGlyAsnLeuAspArgValLeuGluLeu                                    455055                                                                        CTTCGTTCGGGCACCGATATCAACACATGCAATGCGAATGGC260                                 LeuArgSerGlyThrAspIleAsnThrCysAsnAlaAsnGly                                    606570                                                                        CTTAATGCATTGCATCTGGCCTCCAAAGAAGGTCATCATGAA302                                 LeuAsnAlaLeuHisLeuAlaSerLysGluGlyHisHisGlu                                    7580                                                                          GTGGTCCGCGAACTTCTGAAAAGAAAAGCAGATGTTGATGCT344                                 ValValArgGluLeuLeuLysArgLysAlaAspValAspAla                                    859095                                                                        GCCACTAGAAAGGGTAACACAGCGTTACATATAGCATCATTG386                                 AlaThrArgLysGlyAsnThrAlaLeuHisIleAlaSerLeu                                    100105110                                                                     GCAGGACAAGAACTAATCGTCACAGTACTTGTTGAAAATGGT428                                 AlaGlyGlnGluLeuIleValThrValLeuValGluAsnGly                                    115120125                                                                     GCTAATGTTAACGTACAATCACTAAACGGTTTTACACCACTT470                                 AlaAsnValAsnValGlnSerLeuAsnGlyPheThrProLeu                                    130135140                                                                     TACATGGCTGCACAAGAAAATCACGAATCTGTTGTACGCTAT512                                 TyrMetAlaAlaGlnGluAsnHisGluSerValValArgTyr                                    145150                                                                        CTTCTTGCCCACAATGCCAATCAAGCTTTAAGTACAGAAGAC554                                 LeuLeuAlaHisAsnAlaAsnGlnAlaLeuSerThrGluAsp                                    155160165                                                                     GGTTTTACGCCACTGGCAGTTGCCTTGCAACAAGGTCACGAT596                                 GlyPheThrProLeuAlaValAlaLeuGlnGlnGlyHisAsp                                    170175180                                                                     CGTGTGGTCGCTGTTTTGCTTGAAAATGACACGCGCGGGAAA638                                 ArgValValAlaValLeuLeuGluAsnAspThrArgGlyLys                                    185190195                                                                     GTGCGCTTGCCAGCACTGCATATTGCTGCTAAAAAAGATGAT680                                 ValArgLeuProAlaLeuHisIleAlaAlaLysLysAspAsp                                    200205210                                                                     ACGAAAGCAGCTACGCTATTACTTCAAAATGAGCATAACTCG722                                 ThrLysAlaAlaThrLeuLeuLeuGlnAsnGluHisAsnSer                                    215220                                                                        GATGTGACTTCGAAAAGCGGCTTTACTCCGCTTCATATCGCC764                                 AspValThrSerLysSerGlyPheThrProLeuHisIleAla                                    225230235                                                                     GCTCACTATGGAAATGAGAACGTAGCACAACTGCTACTCGAA806                                 AlaHisTyrGlyAsnGluAsnValAlaGlnLeuLeuLeuGlu                                    240245250                                                                     AAGGGAGCCAATGTGAATTACCAAGCGAGACATAACATAAGT848                                 LysGlyAlaAsnValAsnTyrGlnAlaArgHisAsnIleSer                                    255260265                                                                     CCGTTACACGTTGCAACAAAATGGGGTCGTACAAACATGGTT890                                 ProLeuHisValAlaThrLysTrpGlyArgThrAsnMetVal                                    270275280                                                                     TCGTTATTGTTGGCTCATGGGGCCGTAATTGACTGTCGCACA932                                 SerLeuLeuLeuAlaHisGlyAlaValIleAspCysArgThr                                    285290                                                                        CGTGATTTACTAACACCATTACACTGTGCTTCTCGTTCAGGT974                                 ArgAspLeuLeuThrProLeuHisCysAlaSerArgSerGly                                    295300305                                                                     CATGATCAAGTTGTTGATTTGTTGCTTGAAAAAGGAGCTCCA1016                                HisAspGlnValValAspLeuLeuLeuGluLysGlyAlaPro                                    310315320                                                                     ATCAGTGCTAAGACAAAAAATGGTTTGGCTCCCTTACATATG1058                                IleSerAlaLysThrLysAsnGlyLeuAlaProLeuHisMet                                    325330335                                                                     GCAGCACAGGTGGATGATGTTACTGTTGACTATCTCACTCCT1100                                AlaAlaGlnValAspAspValThrValAspTyrLeuThrPro                                    340345350                                                                     CTTCATGTGGCTGCTCATTGCGGACATGTCCGTGTCGCTAAA1142                                LeuHisValAlaAlaHisCysGlyHisValArgValAlaLys                                    355360                                                                        CTTTTGCTGGATCGTAATGCTGACCCGAATGCTCGAGCTCTC1184                                LeuLeuLeuAspArgAsnAlaAspProAsnAlaArgAlaLeu                                    365370375                                                                     AATGGCTTCACACCGCTGCATATCGCTTGCAAAAAAAATCGC1226                                AsnGlyPheThrProLeuHisIleAlaCysLysLysAsnArg                                    380385390                                                                     ATTAAAATTGTCGAACTGCTACTGAAATACCACGCTGCAATC1268                                IleLysIleValGluLeuLeuLeuLysTyrHisAlaAlaIle                                    395400405                                                                     GAAGCAACTACTGAATCCGGTCTCTCACCGCTGCATGTCGCT1310                                GluAlaThrThrGluSerGlyLeuSerProLeuHisValAla                                    410415420                                                                     GCTTTTATGGGTGCTATAAACATTGTCATCTATTTACTACAA1352                                AlaPheMetGlyAlaIleAsnIleValIleTyrLeuLeuGln                                    425430                                                                        CAAGGTGCTAATGCAGATGTGGCTACAGTACGCGGTGAAACG1394                                GlnGlyAlaAsnAlaAspValAlaThrValArgGlyGluThr                                    435440445                                                                     CCTCTTCATTTAGCTGCACGAGCAAACCAAACGGACATTGTT1436                                ProLeuHisLeuAlaAlaArgAlaAsnGlnThrAspIleVal                                    450455460                                                                     CGTGTTTTGGTGCGTAATGGAGCACAGGTGGATGCTGCTGCT1478                                ArgValLeuValArgAsnGlyAlaGlnValAspAlaAlaAla                                    465470475                                                                     CGTGAACTACAAACTCCACTGCACATTGCATCACGTCTTGGT1520                                ArgGluLeuGlnThrProLeuHisIleAlaSerArgLeuGly                                    480485490                                                                     AATACCGACATCGTCATTTTGTTGCTGCAGGCTAATGCATCA1562                                AsnThrAspIleValIleLeuLeuLeuGlnAlaAsnAlaSer                                    495500                                                                        CCAAATGCTGCCACAAGAGATCTTTATACTCCTCTTCATATT1604                                ProAsnAlaAlaThrArgAspLeuTyrThrProLeuHisIle                                    505510515                                                                     GCTGCCAAGGAGGGGCAAGAGGAAGTGGCAGCAATATTGATG1646                                AlaAlaLysGluGlyGlnGluGluValAlaAlaIleLeuMet                                    520525530                                                                     GATCATGGAACCGACAAGACACTGCTCACGAAAAAGGGTTTT1688                                AspHisGlyThrAspLysThrLeuLeuThrLysLysGlyPhe                                    535540545                                                                     ACGCCGTTGCATTTAGCTGCTAAGTATGGCAATTTGCCGGTC1730                                ThrProLeuHisLeuAlaAlaLysTyrGlyAsnLeuProVal                                    550555560                                                                     GCGAAATCATTGCTAGAACGAGGAACACCGGTTGACATTGAA1772                                AlaLysSerLeuLeuGluArgGlyThrProValAspIleGlu                                    565570                                                                        GGCAAGAATCAGGTAACACCTCTGCATGTAGCGGCACATTAC1814                                GlyLysAsnGlnValThrProLeuHisValAlaAlaHisTyr                                    575580585                                                                     AATAACGACAAGGTAGCATTGTTACTTCTAGAAAATGGTGCT1856                                AsnAsnAspLysValAlaLeuLeuLeuLeuGluAsnGlyAla                                    590595600                                                                     TCTGCACATGCCGCTGCCAAGAATGGGTACACTCCTTTACAT1898                                SerAlaHisAlaAlaAlaLysAsnGlyTyrThrProLeuHis                                    605610615                                                                     ATTGCCGCGAAGAAGAATCAGATGGATATTGCTAGCACTCTC1940                                IleAlaAlaLysLysAsnGlnMetAspIleAlaSerThrLeu                                    620625630                                                                     CTTCATTATAAGGCAAATGCGAATGCTGAAAGCAAAGCTGGC1982                                LeuHisTyrLysAlaAsnAlaAsnAlaGluSerLysAlaGly                                    635640                                                                        TTTACACCACTTCATCTTGCCGCCCAGGAGGGCCATCGCGAA2024                                PheThrProLeuHisLeuAlaAlaGlnGluGlyHisArgGlu                                    645650655                                                                     ATGGCTGCGTTATTAATTGAAAATGGAGCAAAAGTTGGAGCT2066                                MetAlaAlaLeuLeuIleGluAsnGlyAlaLysValGlyAla                                    660665670                                                                     CAGGCAAGGAATGGCTTGACACCAATGCATTTATGTGCACAG2108                                GlnAlaArgAsnGlyLeuThrProMetHisLeuCysAlaGln                                    675680685                                                                     GAGGATCGTGTGAGCGTAGCAGAAGAACTAGTGAAAGAAAAC2150                                GluAspArgValSerValAlaGluGluLeuValLysGluAsn                                    690695700                                                                     GCAGCCATTGATCCCAAAACGAAAGCAGGATATACGCCGTTA2192                                AlaAlaIleAspProLysThrLysAlaGlyTyrThrProLeu                                    705710                                                                        CATGTTGCTTGCCATTTTGGACAAATAAACATGGTCCGTTTC2234                                HisValAlaCysHisPheGlyGlnIleAsnMetValArgPhe                                    715720725                                                                     TTGATTGAGCATGGCGCACGAGTTTCAGTTATTACTCGTGCT2276                                LeuIleGluHisGlyAlaArgValSerValIleThrArgAla                                    730735740                                                                     TCCTATACTCCTCTGCATCAAGCTGCTCAGCAAGGGCATAAC2318                                SerTyrThrProLeuHisGlnAlaAlaGlnGlnGlyHisAsn                                    745750755                                                                     AGTGTTGTACGTTACTTGTTGGAACATGGTGCAAGTCCAAAT2360                                SerValValArgTyrLeuLeuGluHisGlyAlaSerProAsn                                    760765770                                                                     GTTCATACATCGACAGGACAAACTCCATTATCGATTGCTGAA2402                                ValHisThrSerThrGlyGlnThrProLeuSerIleAlaGlu                                    775780                                                                        CGTCTAGGGTATGTATCCGTGGTTGAAGCGCTTAAAACAATT2444                                ArgLeuGlyTyrValSerValValGluAlaLeuLysThrIle                                    785790795                                                                     ACCGAGACTACTGTGATAACGGAGACCACAACCGTTACTGAA2486                                ThrGluThrThrValIleThrGluThrThrThrValThrGlu                                    800805810                                                                     GAAAGATATAAACCTCAGAATCCCGAAGCAATGAATGAAACC2528                                GluArgTyrLysProGlnAsnProGluAlaMetAsnGluThr                                    815820825                                                                     ATGTTTTCCGATTCCGAAGATGAAGGTGAAGATAATCAGATC2570                                MetPheSerAspSerGluAspGluGlyGluAspAsnGlnIle                                    830835840                                                                     ACAGCCAATGCTCATGCTCATGATTTCTCAGAAAGCCTCACA2612                                ThrAlaAsnAlaHisAlaHisAspPheSerGluSerLeuThr                                    845850                                                                        AAAGGTTTGCACGATTCAACTGGTGTACATTTGATTCATGCC2654                                LysGlyLeuHisAspSerThrGlyValHisLeuIleHisAla                                    855860865                                                                     ACAGAACCGACATTGTCACGAAGTCCGGAAGTGGAAGGTACG2696                                ThrGluProThrLeuSerArgSerProGluValGluGlyThr                                    870875880                                                                     GATGGCGATTTGGATGCCTTAATTCGTAAAGCACAACATGAA2738                                AspGlyAspLeuAspAlaLeuIleArgLysAlaGlnHisGlu                                    885890895                                                                     CCAATTACTACAGCGATGGCCGATCCTTCCTTAGATGCATCG2780                                ProIleThrThrAlaMetAlaAspProSerLeuAspAlaSer                                    900905910                                                                     CTTCCTGACAATGTTACGATAATGAGAACTACCATGCAACCT2822                                LeuProAspAsnValThrIleMetArgThrThrMetGlnPro                                    915920                                                                        AGTTTTTTAATTTCGTTTATGGTGGATGCACGTGGAGGAGCA2864                                SerPheLeuIleSerPheMetValAspAlaArgGlyGlyAla                                    925930935                                                                     ATGCGTGGTTGTAGGCATTCCGGTGTCAGAATCATTATACCA2906                                MetArgGlyCysArgHisSerGlyValArgIleIleIlePro                                    940945950                                                                     CCGAGGAAAGCGCCGCAACCTACACGGGTCACATGCAGATAC2948                                ProArgLysAlaProGlnProThrArgValThrCysArgTyr                                    955960965                                                                     CTTGGAAAGGACAAGTTAGCGCATCCACCACCATTAAGTGAA2990                                LeuGlyLysAspLysLeuAlaHisProProProLeuSerGlu                                    970975980                                                                     GGTGAAGCGCTCGCNTCACGTATACTTGAAATGGCACCACAT3032                                GlyGluAlaLeuAlaSerArgIleLeuGluMetAlaProHis                                    985990                                                                        GGAGCAAAATTCTTAGGCCCTGTTATATTGGAAGTACCACAT3074                                GlyAlaLysPheLeuGlyProValIleLeuGluValProHis                                    99510001005                                                                   TTTGCATCACTTCGTGGACGAGAGAGAGAGATTGTCATTTTG3116                                PheAlaSerLeuArgGlyArgGluArgGluIleValIleLeu                                    101010151020                                                                  CGTTCTGATGATGGGCAGCATTGGAAAGAGCATCAGCTTGAA3158                                ArgSerAspAspGlyGlnHisTrpLysGluHisGlnLeuGlu                                    102510301035                                                                  GCAACAGAAGATGCTGTACAAGAGGTGCTCAATGAATCGTTT3200                                AlaThrGluAspAlaValGlnGluValLeuAsnGluSerPhe                                    104010451050                                                                  GATGCAGAAGAGTTGTCGCAACTTGATGATTTGCATACATCA3242                                AspAlaGluGluLeuSerGlnLeuAspAspLeuHisThrSer                                    10551060                                                                      CGGATTACGCGTATCCTGACCAATGATTTCCCAATGTATTTC3284                                ArgIleThrArgIleLeuThrAsnAspPheProMetTyrPhe                                    106510701075                                                                  GCGGTCGTTACTCGTGTGCGGCAAGAAGTGCACTGTGTTGGT3326                                AlaValValThrArgValArgGlnGluValHisCysValGly                                    108010851090                                                                  CCAGAAGGTGGTGTAATACTCTCTTCAGTTGTTCCTCATGTG3368                                ProGluGlyGlyValIleLeuSerSerValValProHisVal                                    109511001105                                                                  CAGGCCATATTTCCGGATGGTTCCTTGACTAAGACGATCAAA3410                                GlnAlaIlePheProAspGlySerLeuThrLysThrIleLys                                    111011151120                                                                  GTATCTGTGCAAGCCCAGCCAGTTCCACAAGAGATAGTCACT3452                                ValSerValGlnAlaGlnProValProGlnGluIleValThr                                    11251130                                                                      CGTTTACATGGGAATAGAGTCGCTGTTTCTCCAATTGTAACT3494                                ArgLeuHisGlyAsnArgValAlaValSerProIleValThr                                    113511401145                                                                  GTTGAACCGCGTCGTCGCAAATTCCATAAGCCCATAACGCTG3536                                ValGluProArgArgArgLysPheHisLysProIleThrLeu                                    115011551160                                                                  TGCATACCATTGCCACAAAGCTCAAATAAAGGAATGTTAACA3578                                CysIleProLeuProGlnSerSerAsnLysGlyMetLeuThr                                    116511701175                                                                  CAATATAGTGGCCAACCAGGACAGGAACCACCGACGCTGCGT3620                                GlnTyrSerGlyGlnProGlyGlnGluProProThrLeuArg                                    118011851190                                                                  TTACTCTGCAGTAAAACTGGAGGTTCTTCTCCTGCACAGTGG3662                                LeuLeuCysSerLysThrGlyGlySerSerProAlaGlnTrp                                    11951200                                                                      GAAGATATTACTGGAACTACCCAGTTAACATTTACTGGTGAG3704                                GluAspIleThrGlyThrThrGlnLeuThrPheThrGlyGlu                                    120512101215                                                                  GACGTTTCATTTACAACTACGGTTTCTGCTCGATTTTGGTTG3746                                AspValSerPheThrThrThrValSerAlaArgPheTrpLeu                                    122012251230                                                                  ATGGATTGCCAAACTCCGCGAGATGCGGCACGAATGGCACAA3788                                MetAspCysGlnThrProArgAspAlaAlaArgMetAlaGln                                    123512401245                                                                  GAAGTTTACAATGAAGCAATTGCAGTTCCTTATATGGCTAAA3830                                GluValTyrAsnGluAlaIleAlaValProTyrMetAlaLys                                    125012551260                                                                  TTTCTTATTTTTGCTCGACGAACTTTTCCTGCCGAAGGACAG3872                                PheLeuIlePheAlaArgArgThrPheProAlaGluGlyGln                                    12651270                                                                      TTGAGATTGTTTTGTATGACTGATGATCGGGAAGATAAAACC3914                                LeuArgLeuPheCysMetThrAspAspArgGluAspLysThr                                    127512801285                                                                  CTGGAAAAACAAGAACGTTTCATTGAAATTGCGAAATCGAAA3956                                LeuGluLysGlnGluArgPheIleGluIleAlaLysSerLys                                    129012951300                                                                  GATGTAGAAGTCTTAAGTGGGCGACATCAGTTTTTGGAATTT3998                                AspValGluValLeuSerGlyArgHisGlnPheLeuGluPhe                                    130513101315                                                                  TCTGGAAATCTTCTTCCAATAACCAAGAGTGGTGACCAACTT4040                                SerGlyAsnLeuLeuProIleThrLysSerGlyAspGlnLeu                                    132013251330                                                                  TCTCTTTATTTTCTACCATTCCAAGAAAATCGTCTTGCTTTC4082                                SerLeuTyrPheLeuProPheGlnGluAsnArgLeuAlaPhe                                    13351340                                                                      ATGGTAAAGATACGCACTCACACGGACAACGAAACTGCAGCT4124                                MetValLysIleArgThrHisThrAspAsnGluThrAlaAla                                    134513501355                                                                  GATGGCCGGATAGTATTTATGAAAGAACCAAAATTGAGAGCC4166                                AspGlyArgIleValPheMetLysGluProLysLeuArgAla                                    136013651370                                                                  GAAAATTTACCTCCGCAGACGCCAGTGTGTACTCTTGCAATC4208                                GluAsnLeuProProGlnThrProValCysThrLeuAlaIle                                    137513801385                                                                  ACTCTTCCGGAATACACTGGGCCGGAGCCGATGGTTTCCAAA4250                                ThrLeuProGluTyrThrGlyProGluProMetValSerLys                                    139013951400                                                                  AAACTCTTCTATTCGGAAGCTTCTTTGACTGAGAAATACGTT4292                                LysLeuPheTyrSerGluAlaSerLeuThrGluLysTyrVal                                    14051410                                                                      GGAGCTTTCCATGAAACTGCTGAACCTGATAACTTGCCACTA4334                                GlyAlaPheHisGluThrAlaGluProAspAsnLeuProLeu                                    141514201425                                                                  GCACATGTTGCACTATTAATTGGCGCTGATTGGCATCGGTTA4376                                AlaHisValAlaLeuLeuIleGlyAlaAspTrpHisArgLeu                                    143014351440                                                                  GCTCGAGCGCTTGAAGTACCTGATATTGATATACGACAAGTT4418                                AlaArgAlaLeuGluValProAspIleAspIleArgGlnVal                                    144514501455                                                                  CGACATCAACTAGTTGGTCTTGAAGCAGTCACTATTCTACGT4460                                ArgHisGlnLeuValGlyLeuGluAlaValThrIleLeuArg                                    146014651470                                                                  ATTTGGATATTTTTGAAGAAAGAACAAGCTACGCCCGTTGCT4502                                IleTrpIlePheLeuLysLysGluGlnAlaThrProValAla                                    14751480                                                                      TTGCGATCAGCATTGCAGCGAATAGGACGTGATGATGTTGTA4544                                LeuArgSerAlaLeuGlnArgIleGlyArgAspAspValVal                                    148514901495                                                                  CGAGAAATGGATCGAGCTGAAAAGCTAGATGGTTTAGAAGGA4586                                ArgGluMetAspArgAlaGluLysLeuAspGlyLeuGluGly                                    150015051510                                                                  ACACCTGTATCGCATATTTCTGGACCCTCAATAACTCTGTCA4628                                ThrProValSerHisIleSerGlyProSerIleThrLeuSer                                    151515201525                                                                  TCTACTTTGCTAGAGGTAGCAGGCGACAGACGTCGTCACGCC4670                                SerThrLeuLeuGluValAlaGlyAspArgArgArgHisAla                                    153015351540                                                                  GAGGTAACAATGGCGCAACAGCGATTGGCACAAGAACCGTTT4712                                GluValThrMetAlaGlnGlnArgLeuAlaGlnGluProPhe                                    15451550                                                                      TTTCAGCAAGTAGGGTATAATGGGACACCTGGAGATCCAGAA4754                                PheGlnGlnValGlyTyrAsnGlyThrProGlyAspProGlu                                    155515601565                                                                  GAACCCAAAGAACAGTCATTCCACGAAGAGGAAGAGGAAGTT4796                                GluProLysGluGlnSerPheHisGluGluGluGluGluVal                                    157015751580                                                                  GCAGTTTCAGAAATTCGAACAGTTGTGCGCACTGAACGACAT4838                                AlaValSerGluIleArgThrValValArgThrGluArgHis                                    158515901595                                                                  GTGCATGATTCGGAAAATGGTCCTATTGTGGAAGAGCGTACA4880                                ValHisAspSerGluAsnGlyProIleValGluGluArgThr                                    160016051610                                                                  ATAACAACTACGTATGAGGATGATGTTGCTGTAAACGAAGAA4922                                IleThrThrThrTyrGluAspAspValAlaValAsnGluGlu                                    16151620                                                                      GAAATTGTTGACAAAATAGTGCCTCTCAACGAAGAGGAGCAA4964                                GluIleValAspLysIleValProLeuAsnGluGluGluGln                                    162516301635                                                                  GAAAAATGGGATCGAATGGTTCGAGAAGTGGAAATGAATTTT5006                                GluLysTrpAspArgMetValArgGluValGluMetAsnPhe                                    164016451650                                                                  GAGCAACAAGAAACATCAAAAGAAGGAACGTTTGGTTGTCAG5048                                GluGlnGlnGluThrSerLysGluGlyThrPheGlyCysGln                                    165516601665                                                                  ACAACACATGAGAAAGAAAAAGATGATGATGGTGGCAGTCTG5090                                ThrThrHisGluLysGluLysAspAspAspGlyGlySerLeu                                    167016751680                                                                  AAGACGACAATGAAAGATAGTCACGTAAGGCAGATTTTCTTC5132                                LysThrThrMetLysAspSerHisValArgGlnIlePhePhe                                    16851690                                                                      GATGGAGGTGAGACATCCGCTAATGAAACAGGATTAAGTAGC5174                                AspGlyGlyGluThrSerAlaAsnGluThrGlyLeuSerSer                                    169517001705                                                                  GGGGATGCAGACACTATTATGACTCCAACGACAAAGGAGGAT5216                                GlyAspAlaAspThrIleMetThrProThrThrLysGluAsp                                    171017151720                                                                  AATCATGTTATAGACGTAATGGAGGAAAGGCGAACTGATGAA5258                                AsnHisValIleAspValMetGluGluArgArgThrAspGlu                                    172517301735                                                                  GAGGCCAAAGGGCAAAGCGTTCATGAATAATCTGGATCCA5298                                  GluAlaLysGlyGlnSerValHisGlu                                                   17401745                                                                      CAAATTGATTTAAATCGCAATCTCGCACATGCCTATGTTGCTAATATTTA5348                        ATGAAATTTTTCAAAGCAATAATTTGAATGCTGTTTGGGCTTCCCATATT5398                        GTTAAAGCGTTTTCCATCGTCCATTCACTTTTTGTTTTTGCTGTAGTCTG5448                        TAACTGCTACTCTTGATAAATTTGCTCCAGTAAAAAAAAAAAAAAAAAAA5498                        AAAAA5503                                                                     (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 1745 amino acids                                                  (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      MetSerAsnProIleValGluGlySerGlyTrpProAlaGlu                                    1510                                                                          ProLysAspSerGlnHisGlnGlnGlnIleProAspAspAsn                                    152025                                                                        SerGlnHisSerAsnLysGlyGluSerSerAlaSerPheLeu                                    303540                                                                        ArgAlaAlaArgAlaGlyAsnLeuAspArgValLeuGluLeu                                    455055                                                                        LeuArgSerGlyThrAspIleAsnThrCysAsnAlaAsnGly                                    606570                                                                        LeuAsnAlaLeuHisLeuAlaSerLysGluGlyHisHisGlu                                    7580                                                                          ValValArgGluLeuLeuLysArgLysAlaAspValAspAla                                    859095                                                                        AlaThrArgLysGlyAsnThrAlaLeuHisIleAlaSerLeu                                    100105110                                                                     AlaGlyGlnGluLeuIleValThrValLeuValGluAsnGly                                    115120125                                                                     AlaAsnValAsnValGlnSerLeuAsnGlyPheThrProLeu                                    130135140                                                                     TyrMetAlaAlaGlnGluAsnHisGluSerValValArgTyr                                    145150                                                                        LeuLeuAlaHisAsnAlaAsnGlnAlaLeuSerThrGluAsp                                    155160165                                                                     GlyPheThrProLeuAlaValAlaLeuGlnGlnGlyHisAsp                                    170175180                                                                     ArgValValAlaValLeuLeuGluAsnAspThrArgGlyLys                                    185190195                                                                     ValArgLeuProAlaLeuHisIleAlaAlaLysLysAspAsp                                    200205210                                                                     ThrLysAlaAlaThrLeuLeuLeuGlnAsnGluHisAsnSer                                    215220                                                                        AspValThrSerLysSerGlyPheThrProLeuHisIleAla                                    225230235                                                                     AlaHisTyrGlyAsnGluAsnValAlaGlnLeuLeuLeuGlu                                    240245250                                                                     LysGlyAlaAsnValAsnTyrGlnAlaArgHisAsnIleSer                                    255260265                                                                     ProLeuHisValAlaThrLysTrpGlyArgThrAsnMetVal                                    270275280                                                                     SerLeuLeuLeuAlaHisGlyAlaValIleAspCysArgThr                                    285290                                                                        ArgAspLeuLeuThrProLeuHisCysAlaSerArgSerGly                                    295300305                                                                     HisAspGlnValValAspLeuLeuLeuGluLysGlyAlaPro                                    310315320                                                                     IleSerAlaLysThrLysAsnGlyLeuAlaProLeuHisMet                                    325330335                                                                     AlaAlaGlnValAspAspValThrValAspTyrLeuThrPro                                    340345350                                                                     LeuHisValAlaAlaHisCysGlyHisValArgValAlaLys                                    355360                                                                        LeuLeuLeuAspArgAsnAlaAspProAsnAlaArgAlaLeu                                    365370375                                                                     AsnGlyPheThrProLeuHisIleAlaCysLysLysAsnArg                                    380385390                                                                     IleLysIleValGluLeuLeuLeuLysTyrHisAlaAlaIle                                    395400405                                                                     GluAlaThrThrGluSerGlyLeuSerProLeuHisValAla                                    410415420                                                                     AlaPheMetGlyAlaIleAsnIleValIleTyrLeuLeuGln                                    425430                                                                        GlnGlyAlaAsnAlaAspValAlaThrValArgGlyGluThr                                    435440445                                                                     ProLeuHisLeuAlaAlaArgAlaAsnGlnThrAspIleVal                                    450455460                                                                     ArgValLeuValArgAsnGlyAlaGlnValAspAlaAlaAla                                    465470475                                                                     ArgGluLeuGlnThrProLeuHisIleAlaSerArgLeuGly                                    480485490                                                                     AsnThrAspIleValIleLeuLeuLeuGlnAlaAsnAlaSer                                    495500                                                                        ProAsnAlaAlaThrArgAspLeuTyrThrProLeuHisIle                                    505510515                                                                     AlaAlaLysGluGlyGlnGluGluValAlaAlaIleLeuMet                                    520525530                                                                     AspHisGlyThrAspLysThrLeuLeuThrLysLysGlyPhe                                    535540545                                                                     ThrProLeuHisLeuAlaAlaLysTyrGlyAsnLeuProVal                                    550555560                                                                     AlaLysSerLeuLeuGluArgGlyThrProValAspIleGlu                                    565570                                                                        GlyLysAsnGlnValThrProLeuHisValAlaAlaHisTyr                                    575580585                                                                     AsnAsnAspLysValAlaLeuLeuLeuLeuGluAsnGlyAla                                    590595600                                                                     SerAlaHisAlaAlaAlaLysAsnGlyTyrThrProLeuHis                                    605610615                                                                     IleAlaAlaLysLysAsnGlnMetAspIleAlaSerThrLeu                                    620625630                                                                     LeuHisTyrLysAlaAsnAlaAsnAlaGluSerLysAlaGly                                    635640                                                                        PheThrProLeuHisLeuAlaAlaGlnGluGlyHisArgGlu                                    645650655                                                                     MetAlaAlaLeuLeuIleGluAsnGlyAlaLysValGlyAla                                    660665670                                                                     GlnAlaArgAsnGlyLeuThrProMetHisLeuCysAlaGln                                    675680685                                                                     GluAspArgValSerValAlaGluGluLeuValLysGluAsn                                    690695700                                                                     AlaAlaIleAspProLysThrLysAlaGlyTyrThrProLeu                                    705710                                                                        HisValAlaCysHisPheGlyGlnIleAsnMetValArgPhe                                    715720725                                                                     LeuIleGluHisGlyAlaArgValSerValIleThrArgAla                                    730735740                                                                     SerTyrThrProLeuHisGlnAlaAlaGlnGlnGlyHisAsn                                    745750755                                                                     SerValValArgTyrLeuLeuGluHisGlyAlaSerProAsn                                    760765770                                                                     ValHisThrSerThrGlyGlnThrProLeuSerIleAlaGlu                                    775780                                                                        ArgLeuGlyTyrValSerValValGluAlaLeuLysThrIle                                    785790795                                                                     ThrGluThrThrValIleThrGluThrThrThrValThrGlu                                    800805810                                                                     GluArgTyrLysProGlnAsnProGluAlaMetAsnGluThr                                    815820825                                                                     MetPheSerAspSerGluAspGluGlyGluAspAsnGlnIle                                    830835840                                                                     ThrAlaAsnAlaHisAlaHisAspPheSerGluSerLeuThr                                    845850                                                                        LysGlyLeuHisAspSerThrGlyValHisLeuIleHisAla                                    855860865                                                                     ThrGluProThrLeuSerArgSerProGluValGluGlyThr                                    870875880                                                                     AspGlyAspLeuAspAlaLeuIleArgLysAlaGlnHisGlu                                    885890895                                                                     ProIleThrThrAlaMetAlaAspProSerLeuAspAlaSer                                    900905910                                                                     LeuProAspAsnValThrIleMetArgThrThrMetGlnPro                                    915920                                                                        SerPheLeuIleSerPheMetValAspAlaArgGlyGlyAla                                    925930935                                                                     MetArgGlyCysArgHisSerGlyValArgIleIleIlePro                                    940945950                                                                     ProArgLysAlaProGlnProThrArgValThrCysArgTyr                                    955960965                                                                     LeuGlyLysAspLysLeuAlaHisProProProLeuSerGlu                                    970975980                                                                     GlyGluAlaLeuAlaSerArgIleLeuGluMetAlaProHis                                    985990                                                                        GlyAlaLysPheLeuGlyProValIleLeuGluValProHis                                    99510001005                                                                   PheAlaSerLeuArgGlyArgGluArgGluIleValIleLeu                                    101010151020                                                                  ArgSerAspAspGlyGlnHisTrpLysGluHisGlnLeuGlu                                    102510301035                                                                  AlaThrGluAspAlaValGlnGluValLeuAsnGluSerPhe                                    104010451050                                                                  AspAlaGluGluLeuSerGlnLeuAspAspLeuHisThrSer                                    10551060                                                                      ArgIleThrArgIleLeuThrAsnAspPheProMetTyrPhe                                    106510701075                                                                  AlaValValThrArgValArgGlnGluValHisCysValGly                                    108010851090                                                                  ProGluGlyGlyValIleLeuSerSerValValProHisVal                                    109511001105                                                                  GlnAlaIlePheProAspGlySerLeuThrLysThrIleLys                                    111011151120                                                                  ValSerValGlnAlaGlnProValProGlnGluIleValThr                                    11251130                                                                      ArgLeuHisGlyAsnArgValAlaValSerProIleValThr                                    113511401145                                                                  ValGluProArgArgArgLysPheHisLysProIleThrLeu                                    115011551160                                                                  CysIleProLeuProGlnSerSerAsnLysGlyMetLeuThr                                    116511701175                                                                  GlnTyrSerGlyGlnProGlyGlnGluProProThrLeuArg                                    118011851190                                                                  LeuLeuCysSerLysThrGlyGlySerSerProAlaGlnTrp                                    11951200                                                                      GluAspIleThrGlyThrThrGlnLeuThrPheThrGlyGlu                                    120512101215                                                                  AspValSerPheThrThrThrValSerAlaArgPheTrpLeu                                    122012251230                                                                  MetAspCysGlnThrProArgAspAlaAlaArgMetAlaGln                                    123512401245                                                                  GluValTyrAsnGluAlaIleAlaValProTyrMetAlaLys                                    125012551260                                                                  PheLeuIlePheAlaArgArgThrPheProAlaGluGlyGln                                    12651270                                                                      LeuArgLeuPheCysMetThrAspAspArgGluAspLysThr                                    127512801285                                                                  LeuGluLysGlnGluArgPheIleGluIleAlaLysSerLys                                    129012951300                                                                  AspValGluValLeuSerGlyArgHisGlnPheLeuGluPhe                                    130513101315                                                                  SerGlyAsnLeuLeuProIleThrLysSerGlyAspGlnLeu                                    132013251330                                                                  SerLeuTyrPheLeuProPheGlnGluAsnArgLeuAlaPhe                                    13351340                                                                      MetValLysIleArgThrHisThrAspAsnGluThrAlaAla                                    134513501355                                                                  AspGlyArgIleValPheMetLysGluProLysLeuArgAla                                    136013651370                                                                  GluAsnLeuProProGlnThrProValCysThrLeuAlaIle                                    137513801385                                                                  ThrLeuProGluTyrThrGlyProGluProMetValSerLys                                    139013951400                                                                  LysLeuPheTyrSerGluAlaSerLeuThrGluLysTyrVal                                    14051410                                                                      GlyAlaPheHisGluThrAlaGluProAspAsnLeuProLeu                                    141514201425                                                                  AlaHisValAlaLeuLeuIleGlyAlaAspTrpHisArgLeu                                    143014351440                                                                  AlaArgAlaLeuGluValProAspIleAspIleArgGlnVal                                    144514501455                                                                  ArgHisGlnLeuValGlyLeuGluAlaValThrIleLeuArg                                    146014651470                                                                  IleTrpIlePheLeuLysLysGluGlnAlaThrProValAla                                    14751480                                                                      LeuArgSerAlaLeuGlnArgIleGlyArgAspAspValVal                                    148514901495                                                                  ArgGluMetAspArgAlaGluLysLeuAspGlyLeuGluGly                                    150015051510                                                                  ThrProValSerHisIleSerGlyProSerIleThrLeuSer                                    151515201525                                                                  SerThrLeuLeuGluValAlaGlyAspArgArgArgHisAla                                    153015351540                                                                  GluValThrMetAlaGlnGlnArgLeuAlaGlnGluProPhe                                    15451550                                                                      PheGlnGlnValGlyTyrAsnGlyThrProGlyAspProGlu                                    155515601565                                                                  GluProLysGluGlnSerPheHisGluGluGluGluGluVal                                    157015751580                                                                  AlaValSerGluIleArgThrValValArgThrGluArgHis                                    158515901595                                                                  ValHisAspSerGluAsnGlyProIleValGluGluArgThr                                    160016051610                                                                  IleThrThrThrTyrGluAspAspValAlaValAsnGluGlu                                    16151620                                                                      GluIleValAspLysIleValProLeuAsnGluGluGluGln                                    162516301635                                                                  GluLysTrpAspArgMetValArgGluValGluMetAsnPhe                                    164016451650                                                                  GluGlnGlnGluThrSerLysGluGlyThrPheGlyCysGln                                    165516601665                                                                  ThrThrHisGluLysGluLysAspAspAspGlyGlySerLeu                                    167016751680                                                                  LysThrThrMetLysAspSerHisValArgGlnIlePhePhe                                    16851690                                                                      AspGlyGlyGluThrSerAlaAsnGluThrGlyLeuSerSer                                    169517001705                                                                  GlyAspAlaAspThrIleMetThrProThrThrLysGluAsp                                    171017151720                                                                  AsnHisValIleAspValMetGluGluArgArgThrAspGlu                                    172517301735                                                                  GluAlaLysGlyGlnSerValHisGlu                                                   17401745                                                                      (2) INFORMATION FOR SEQ ID NO:34:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5503 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                      TTTTTTTTTTTTTTTTTTTTTTTTACTGGAGCAAATTTATCAAGAGTAGC50                          AGTTACAGACTACAGCAAAAACAAAAAGTGAATGGACGATGGAAAACGCT100                         TTAACAATATGGGAAGCCCAAACAGCATTCAAATTATTGCTTTGAAAAAT150                         TTCATTAAATATTAGCAACATAGGCATGTGCGAGATTGCGATTTAAATCA200                         ATTTGTGGATCCAGATTATTCATGAACGCTTTGCCCTTTGGCCTCTTCAT250                         CAGTTCGCCTTTCCTCCATTACGTCTATAACATGATTATCCTCCTTTGTC300                         GTTGGAGTCATAATAGTGTCTGCATCCCCGCTACTTAATCCTGTTTCATT350                         AGCGGATGTCTCACCTCCATCGAAGAAAATCTGCCTTACGTGACTATCTT400                         TCATTGTCGTCTTCAGACTGCCACCATCATCATCTTTTTCTTTCTCATGT450                         GTTGTCTGACAACCAAACGTTCCTTCTTTTGATGTTTCTTGTTGCTCAAA500                         ATTCATTTCCACTTCTCGAACCATTCGATCCCATTTTTCTTGCTCCTCTT550                         CGTTGAGAGGCACTATTTTGTCAACAATTTCTTCTTCGTTTACAGCAACA600                         TCATCCTCATACGTAGTTGTTATTGTACGCTCTTCCACAATAGGACCATT650                         TTCCGAATCATGCACATGTCGTTCAGTGCGCACAACTGTTCGAATTTCTG700                         AAACTGCAACTTCCTCTTCCTCTTCGTGGAATGACTGTTCTTTGGGTTCT750                         TCTGGATCTCCAGGTGTCCCATTATACCCTACTTGCTGAAAAAACGGTTC800                         TTGTGCCAATCGCTGTTGCGCCATTGTTACCTCGGCGTGACGACGTCTGT850                         CGCCTGCTACCTCTAGCAAAGTAGATGACAGAGTTATTGAGGGTCCAGAA900                         ATATGCGATACAGGTGTTCCTTCTAAACCATCTAGCTTTTCAGCTCGATC950                         CATTTCTCGTACAACATCATCACGTCCTATTCGCTGCAATGCTGATCGCA1000                        AAGCAACGGGCGTAGCTTGTTCTTTCTTCAAAAATATCCAAATACGTAGA1050                        ATAGTGACTGCTTCAAGACCAACTAGTTGATGTCGAACTTGTCGTATATC1100                        AATATCAGGTACTTCAAGCGCTCGAGCTAACCGATGCCAATCAGCGCCAA1150                        TTAATAGTGCAACATGTGCTAGTGGCAAGTTATCAGGTTCAGCAGTTTCA1200                        TGGAAAGCTCCAACGTATTTCTCAGTCAAAGAAGCTTCCGAATAGAAGAG1250                        TTTTTTGGAAACCATCGGCTCCGGCCCAGTGTATTCCGGAAGAGTGATTG1300                        CAAGAGTACACACTGGCGTCTGCGGAGGTAAATTTTCGGCTCTCAATTTT1350                        GGTTCTTTCATAAATACTATCCGGCCATCAGCTGCAGTTTCGTTGTCCGT1400                        GTGAGTGCGTATCTTTACCATGAAAGCAAGACGATTTTCTTGGAATGGTA1450                        GAAAATAAAGAGAAAGTTGGTCACCACTCTTGGTTATTGGAAGAAGATTT1500                        CCAGAAAATTCCAAAAACTGATGTCGCCCACTTAAGACTTCTACATCTTT1550                        CGATTTCGCAATTTCAATGAAACGTTCTTGTTTTTCCAGGGTTTTATCTT1600                        CCCGATCATCAGTCATACAAAACAATCTCAACTGTCCTTCGGCAGGAAAA1650                        GTTCGTCGAGCAAAAATAAGAAATTTAGCCATATAAGGAACTGCAATTGC1700                        TTCATTGTAAACTTCTTGTGCCATTCGTGCCGCATCTCGCGGAGTTTGGC1750                        AATCCATCAACCAAAATCGAGCAGAAACCGTAGTTGTAAATGAAACGTCC1800                        TCACCAGTAAATGTTAACTGGGTAGTTCCAGTAATATCTTCCCACTGTGC1850                        AGGAGAAGAACCTCCAGTTTTACTGCAGAGTAAACGCAGCGTCGGTGGTT1900                        CCTGTCCTGGTTGGCCACTATATTGTGTTAACATTCCTTTATTTGAGCTT1950                        TGTGGCAATGGTATGCACAGCGTTATGGGCTTATGGAATTTGCGACGACG2000                        CGGTTCAACAGTTACAATTGGAGAAACAGCGACTCTATTCCCATGTAAAC2050                        GAGTGACTATCTCTTGTGGAACTGGCTGGGCTTGCACAGATACTTTGATC2100                        GTCTTAGTCAAGGAACCATCCGGAAATATGGCCTGCACATGAGGAACAAC2150                        TGAAGAGAGTATTACACCACCTTCTGGACCAACACAGTGCACTTCTTGCC2200                        GCACACGAGTAACGACCGCGAAATACATTGGGAAATCATTGGTCAGGATA2250                        CGCGTAATCCGTGATGTATGCAAATCATCAAGTTGCGACAACTCTTCTGC2300                        ATCAAACGATTCATTGAGCACCTCTTGTACAGCATCTTCTGTTGCTTCAA2350                        GCTGATGCTCTTTCCAATGCTGCCCATCATCAGAACGCAAAATGACAATC2400                        TCTCTCTCTCGTCCACGAAGTGATGCAAAATGTGGTACTTCCAATATAAC2450                        AGGGCCTAAGAATTTTGCTCCATGTGGTGCCATTTCAAGTATACGTGANG2500                        CGAGCGCTTCACCTTCACTTAATGGTGGTGGATGCGCTAACTTGTCCTTT2550                        CCAAGGTATCTGCATGTGACCCGTGTAGGTTGCGGCGCTTTCCTCGGTGG2600                        TATAATGATTCTGACACCGGAATGCCTACAACCACGCATTGCTCCTCCAC2650                        GTGCATCCACCATAAACGAAATTAAAAAACTAGGTTGCATGGTAGTTCTC2700                        ATTATCGTAACATTGTCAGGAAGCGATGCATCTAAGGAAGGATCGGCCAT2750                        CGCTGTAGTAATTGGTTCATGTTGTGCTTTACGAATTAAGGCATCCAAAT2800                        CGCCATCCGTACCTTCCACTTCCGGACTTCGTGACAATGTCGGTTCTGTG2850                        GCATGAATCAAATGTACACCAGTTGAATCGTGCAAACCTTTTGTGAGGCT2900                        TTCTGAGAAATCATGAGCATGAGCATTGGCTGTGATCTGATTATCTTCAC2950                        CTTCATCTTCGGAATCGGAAAACATGGTTTCATTCATTGCTTCGGGATTC3000                        TGAGGTTTATATCTTTCTTCAGTAACGGTTGTGGTCTCCGTTATCACAGT3050                        AGTCTCGGTAATTGTTTTAAGCGCTTCAACCACGGATACATACCCTAGAC3100                        GTTCAGCAATCGATAATGGAGTTTGTCCTGTCGATGTATGAACATTTGGA3150                        CTTGCACCATGTTCCAACAAGTAACGTACAACACTGTTATGCCCTTGCTG3200                        AGCAGCTTGATGCAGAGGAGTATAGGAAGCACGAGTAATAACTGAAACTC3250                        GTGCGCCATGCTCAATCAAGAAACGGACCATGTTTATTTGTCCAAAATGG3300                        CAAGCAACATGTAACGGCGTATATCCTGCTTTCGTTTTGGGATCAATGGC3350                        TGCGTTTTCTTTCACTAGTTCTTCTGCTACGCTCACACGATCCTCCTGTG3400                        CACATAAATGCATTGGTGTCAAGCCATTCCTTGCCTGAGCTCCAACTTTT3450                        GCTCCATTTTCAATTAATAACGCAGCCATTTCGCGATGGCCCTCCTGGGC3500                        GGCAAGATGAAGTGGTGTAAAGCCAGCTTTGCTTTCAGCATTCGCATTTG3550                        CCTTATAATGAAGGAGAGTGCTAGCAATATCCATCTGATTCTTCTTCGCG3600                        GCAATATGTAAAGGAGTGTACCCATTCTTGGCAGCGGCATGTGCAGAAGC3650                        ACCATTTTCTAGAAGTAACAATGCTACCTTGTCGTTATTGTAATGTGCCG3700                        CTACATGCAGAGGTGTTACCTGATTCTTGCCTTCAATGTCAACCGGTGTT3750                        CCTCGTTCTAGCAATGATTTCGCGACCGGCAAATTGCCATACTTAGCAGC3800                        TAAATGCAACGGCGTAAAACCCTTTTTCGTGAGCAGTGTCTTGTCGGTTC3850                        CATGATCCATCAATATTGCTGCCACTTCCTCTTGCCCCTCCTTGGCAGCA3900                        ATATGAAGAGGAGTATAAAGATCTCTTGTGGCAGCATTTGGTGATGCATT3950                        AGCCTGCAGCAACAAAATGACGATGTCGGTATTACCAAGACGTGATGCAA4000                        TGTGCAGTGGAGTTTGTAGTTCACGAGCAGCAGCATCCACCTGTGCTCCA4050                        TTACGCACCAAAACACGAACAATGTCCGTTTGGTTTGCTCGTGCAGCTAA4100                        ATGAAGAGGCGTTTCACCGCGTACTGTAGCCACATCTGCATTAGCACCTT4150                        GTTGTAGTAAATAGATGACAATGTTTATAGCACCCATAAAAGCAGCGACA4200                        TGCAGCGGTGAGAGACCGGATTCAGTAGTTGCTTCGATTGCAGCGTGGTA4250                        TTTCAGTAGCAGTTCGACAATTTTAATGCGATTTTTTTTGCAAGCGATAT4300                        GCAGCGGTGTGAAGCCATTGAGAGCTCGAGCATTCGGGTCAGCATTACGA4350                        TCCAGCAAAAGTTTAGCGACACGGACATGTCCGCAATGAGCAGCCACATG4400                        AAGAGGAGTGAGATAGTCAACAGTAACATCATCCACCTGTGCTGCCATAT4450                        GTAAGGGAGCCAAACCATTTTTTGTCTTAGCACTGATTGGAGCTCCTTTT4500                        TCAAGCAACAAATCAACAACTTGATCATGACCTGAACGAGAAGCACAGTG4550                        TAATGGTGTTAGTAAATCACGTGTGCGACAGTCAATTACGGCCCCATGAG4600                        CCAACAATAACGAAACCATGTTTGTACGACCCCATTTTGTTGCAACGTGT4650                        AACGGACTTATGTTATGTCTCGCTTGGTAATTCACATTGGCTCCCTTTTC4700                        GAGTAGCAGTTGTGCTACGTTCTCATTTCCATAGTGAGCGGCGATATGAA4750                        GCGGAGTAAAGCCGCTTTTCGAAGTCACATCCGAGTTATGCTCATTTTGA4800                        AGTAATAGCGTAGCTGCTTTCGTATCATCTTTTTTAGCAGCAATATGCAG4850                        TGCTGGCAAGCGCACTTTCCCGCGCGTGTCATTTTCAAGCAAAACAGCGA4900                        CCACACGATCGTGACCTTGTTGCAAGGCAACTGCCAGTGGCGTAAAACCG4950                        TCTTCTGTACTTAAAGCTTGATTGGCATTGTGGGCAAGAAGATAGCGTAC5000                        AACAGATTCGTGATTTTCTTGTGCAGCCATGTAAAGTGGTGTAAAACCGT5050                        TTAGTGATTGTACGTTAACATTAGCACCATTTTCAACAAGTACTGTGACG5100                        ATTAGTTCTTGTCCTGCCAATGATGCTATATGTAACGCTGTGTTACCCTT5150                        TCTAGTGGCAGCATCAACATCTGCTTTTCTTTTCAGAAGTTCGCGGACCA5200                        CTTCATGATGACCTTCTTTGGAGGCCAGATGCAATGCATTAAGGCCATTC5250                        GCATTGCATGTGTTGATATCGGTGCCCGAACGAAGTAGTTCAAGTACACG5300                        ATCCAAATTTCCAGCTCTTGCTGCTCGTAAAAAACTTGCACTGCTCTCAC5350                        CTTTGTTGGAATGTTGACTGTTATCATCAGGAATTTGTTGTTGATGTTGT5400                        GAATCTTTTGGTTCTGCGGGCCAGCCACTTCCCTCGACTATAGGATTACT5450                        CATCAATAGTTGAGTTATATCAGTCAGCCGCCTCAAACTTGGGTAATTAA5500                        ACC5503                                                                       (2) INFORMATION FOR SEQ ID NO:35:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5235 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                      ATGAGTAATCCTATAGTCGAGGGAAGTGGCTGGCCCGCAGAACCAAAAGA50                          TTCACAACATCAACAACAAATTCCTGATGATAACAGTCAACATTCCAACA100                         AAGGTGAGAGCAGTGCAAGTTTTTTACGAGCAGCAAGAGCTGGAAATTTG150                         GATCGTGTACTTGAACTACTTCGTTCGGGCACCGATATCAACACATGCAA200                         TGCGAATGGCCTTAATGCATTGCATCTGGCCTCCAAAGAAGGTCATCATG250                         AAGTGGTCCGCGAACTTCTGAAAAGAAAAGCAGATGTTGATGCTGCCACT300                         AGAAAGGGTAACACAGCGTTACATATAGCATCATTGGCAGGACAAGAACT350                         AATCGTCACAGTACTTGTTGAAAATGGTGCTAATGTTAACGTACAATCAC400                         TAAACGGTTTTACACCACTTTACATGGCTGCACAAGAAAATCACGAATCT450                         GTTGTACGCTATCTTCTTGCCCACAATGCCAATCAAGCTTTAAGTACAGA500                         AGACGGTTTTACGCCACTGGCAGTTGCCTTGCAACAAGGTCACGATCGTG550                         TGGTCGCTGTTTTGCTTGAAAATGACACGCGCGGGAAAGTGCGCTTGCCA600                         GCACTGCATATTGCTGCTAAAAAAGATGATACGAAAGCAGCTACGCTATT650                         ACTTCAAAATGAGCATAACTCGGATGTGACTTCGAAAAGCGGCTTTACTC700                         CGCTTCATATCGCCGCTCACTATGGAAATGAGAACGTAGCACAACTGCTA750                         CTCGAAAAGGGAGCCAATGTGAATTACCAAGCGAGACATAACATAAGTCC800                         GTTACACGTTGCAACAAAATGGGGTCGTACAAACATGGTTTCGTTATTGT850                         TGGCTCATGGGGCCGTAATTGACTGTCGCACACGTGATTTACTAACACCA900                         TTACACTGTGCTTCTCGTTCAGGTCATGATCAAGTTGTTGATTTGTTGCT950                         TGAAAAAGGAGCTCCAATCAGTGCTAAGACAAAAAATGGTTTGGCTCCCT1000                        TACATATGGCAGCACAGGTGGATGATGTTACTGTTGACTATCTCACTCCT1050                        CTTCATGTGGCTGCTCATTGCGGACATGTCCGTGTCGCTAAACTTTTGCT1100                        GGATCGTAATGCTGACCCGAATGCTCGAGCTCTCAATGGCTTCACACCGC1150                        TGCATATCGCTTGCAAAAAAAATCGCATTAAAATTGTCGAACTGCTACTG1200                        AAATACCACGCTGCAATCGAAGCAACTACTGAATCCGGTCTCTCACCGCT1250                        GCATGTCGCTGCTTTTATGGGTGCTATAAACATTGTCATCTATTTACTAC1300                        AACAAGGTGCTAATGCAGATGTGGCTACAGTACGCGGTGAAACGCCTCTT1350                        CATTTAGCTGCACGAGCAAACCAAACGGACATTGTTCGTGTTTTGGTGCG1400                        TAATGGAGCACAGGTGGATGCTGCTGCTCGTGAACTACAAACTCCACTGC1450                        ACATTGCATCACGTCTTGGTAATACCGACATCGTCATTTTGTTGCTGCAG1500                        GCTAATGCATCACCAAATGCTGCCACAAGAGATCTTTATACTCCTCTTCA1550                        TATTGCTGCCAAGGAGGGGCAAGAGGAAGTGGCAGCAATATTGATGGATC1600                        ATGGAACCGACAAGACACTGCTCACGAAAAAGGGTTTTACGCCGTTGCAT1650                        TTAGCTGCTAAGTATGGCAATTTGCCGGTCGCGAAATCATTGCTAGAACG1700                        AGGAACACCGGTTGACATTGAAGGCAAGAATCAGGTAACACCTCTGCATG1750                        TAGCGGCACATTACAATAACGACAAGGTAGCATTGTTACTTCTAGAAAAT1800                        GGTGCTTCTGCACATGCCGCTGCCAAGAATGGGTACACTCCTTTACATAT1850                        TGCCGCGAAGAAGAATCAGATGGATATTGCTAGCACTCTCCTTCATTATA1900                        AGGCAAATGCGAATGCTGAAAGCAAAGCTGGCTTTACACCACTTCATCTT1950                        GCCGCCCAGGAGGGCCATCGCGAAATGGCTGCGTTATTAATTGAAAATGG2000                        AGCAAAAGTTGGAGCTCAGGCAAGGAATGGCTTGACACCAATGCATTTAT2050                        GTGCACAGGAGGATCGTGTGAGCGTAGCAGAAGAACTAGTGAAAGAAAAC2100                        GCAGCCATTGATCCCAAAACGAAAGCAGGATATACGCCGTTACATGTTGC2150                        TTGCCATTTTGGACAAATAAACATGGTCCGTTTCTTGATTGAGCATGGCG2200                        CACGAGTTTCAGTTATTACTCGTGCTTCCTATACTCCTCTGCATCAAGCT2250                        GCTCAGCAAGGGCATAACAGTGTTGTACGTTACTTGTTGGAACATGGTGC2300                        AAGTCCAAATGTTCATACATCGACAGGACAAACTCCATTATCGATTGCTG2350                        AACGTCTAGGGTATGTATCCGTGGTTGAAGCGCTTAAAACAATTACCGAG2400                        ACTACTGTGATAACGGAGACCACAACCGTTACTGAAGAAAGATATAAACC2450                        TCAGAATCCCGAAGCAATGAATGAAACCATGTTTTCCGATTCCGAAGATG2500                        AAGGTGAAGATAATCAGATCACAGCCAATGCTCATGCTCATGATTTCTCA2550                        GAAAGCCTCACAAAAGGTTTGCACGATTCAACTGGTGTACATTTGATTCA2600                        TGCCACAGAACCGACATTGTCACGAAGTCCGGAAGTGGAAGGTACGGATG2650                        GCGATTTGGATGCCTTAATTCGTAAAGCACAACATGAACCAATTACTACA2700                        GCGATGGCCGATCCTTCCTTAGATGCATCGCTTCCTGACAATGTTACGAT2750                        AATGAGAACTACCATGCAACCTAGTTTTTTAATTTCGTTTATGGTGGATG2800                        CACGTGGAGGAGCAATGCGTGGTTGTAGGCATTCCGGTGTCAGAATCATT2850                        ATACCACCGAGGAAAGCGCCGCAACCTACACGGGTCACATGCAGATACCT2900                        TGGAAAGGACAAGTTAGCGCATCCACCACCATTAAGTGAAGGTGAAGCGC2950                        TCGCNTCACGTATACTTGAAATGGCACCACATGGAGCAAAATTCTTAGGC3000                        CCTGTTATATTGGAAGTACCACATTTTGCATCACTTCGTGGACGAGAGAG3050                        AGAGATTGTCATTTTGCGTTCTGATGATGGGCAGCATTGGAAAGAGCATC3100                        AGCTTGAAGCAACAGAAGATGCTGTACAAGAGGTGCTCAATGAATCGTTT3150                        GATGCAGAAGAGTTGTCGCAACTTGATGATTTGCATACATCACGGATTAC3200                        GCGTATCCTGACCAATGATTTCCCAATGTATTTCGCGGTCGTTACTCGTG3250                        TGCGGCAAGAAGTGCACTGTGTTGGTCCAGAAGGTGGTGTAATACTCTCT3300                        TCAGTTGTTCCTCATGTGCAGGCCATATTTCCGGATGGTTCCTTGACTAA3350                        GACGATCAAAGTATCTGTGCAAGCCCAGCCAGTTCCACAAGAGATAGTCA3400                        CTCGTTTACATGGGAATAGAGTCGCTGTTTCTCCAATTGTAACTGTTGAA3450                        CCGCGTCGTCGCAAATTCCATAAGCCCATAACGCTGTGCATACCATTGCC3500                        ACAAAGCTCAAATAAAGGAATGTTAACACAATATAGTGGCCAACCAGGAC3550                        AGGAACCACCGACGCTGCGTTTACTCTGCAGTAAAACTGGAGGTTCTTCT3600                        CCTGCACAGTGGGAAGATATTACTGGAACTACCCAGTTAACATTTACTGG3650                        TGAGGACGTTTCATTTACAACTACGGTTTCTGCTCGATTTTGGTTGATGG3700                        ATTGCCAAACTCCGCGAGATGCGGCACGAATGGCACAAGAAGTTTACAAT3750                        GAAGCAATTGCAGTTCCTTATATGGCTAAATTTCTTATTTTTGCTCGACG3800                        AACTTTTCCTGCCGAAGGACAGTTGAGATTGTTTTGTATGACTGATGATC3850                        GGGAAGATAAAACCCTGGAAAAACAAGAACGTTTCATTGAAATTGCGAAA3900                        TCGAAAGATGTAGAAGTCTTAAGTGGGCGACATCAGTTTTTGGAATTTTC3950                        TGGAAATCTTCTTCCAATAACCAAGAGTGGTGACCAACTTTCTCTTTATT4000                        TTCTACCATTCCAAGAAAATCGTCTTGCTTTCATGGTAAAGATACGCACT4050                        CACACGGACAACGAAACTGCAGCTGATGGCCGGATAGTATTTATGAAAGA4100                        ACCAAAATTGAGAGCCGAAAATTTACCTCCGCAGACGCCAGTGTGTACTC4150                        TTGCAATCACTCTTCCGGAATACACTGGGCCGGAGCCGATGGTTTCCAAA4200                        AAACTCTTCTATTCGGAAGCTTCTTTGACTGAGAAATACGTTGGAGCTTT4250                        CCATGAAACTGCTGAACCTGATAACTTGCCACTAGCACATGTTGCACTAT4300                        TAATTGGCGCTGATTGGCATCGGTTAGCTCGAGCGCTTGAAGTACCTGAT4350                        ATTGATATACGACAAGTTCGACATCAACTAGTTGGTCTTGAAGCAGTCAC4400                        TATTCTACGTATTTGGATATTTTTGAAGAAAGAACAAGCTACGCCCGTTG4450                        CTTTGCGATCAGCATTGCAGCGAATAGGACGTGATGATGTTGTACGAGAA4500                        ATGGATCGAGCTGAAAAGCTAGATGGTTTAGAAGGAACACCTGTATCGCA4550                        TATTTCTGGACCCTCAATAACTCTGTCATCTACTTTGCTAGAGGTAGCAG4600                        GCGACAGACGTCGTCACGCCGAGGTAACAATGGCGCAACAGCGATTGGCA4650                        CAAGAACCGTTTTTTCAGCAAGTAGGGTATAATGGGACACCTGGAGATCC4700                        AGAAGAACCCAAAGAACAGTCATTCCACGAAGAGGAAGAGGAAGTTGCAG4750                        TTTCAGAAATTCGAACAGTTGTGCGCACTGAACGACATGTGCATGATTCG4800                        GAAAATGGTCCTATTGTGGAAGAGCGTACAATAACAACTACGTATGAGGA4850                        TGATGTTGCTGTAAACGAAGAAGAAATTGTTGACAAAATAGTGCCTCTCA4900                        ACGAAGAGGAGCAAGAAAAATGGGATCGAATGGTTCGAGAAGTGGAAATG4950                        AATTTTGAGCAACAAGAAACATCAAAAGAAGGAACGTTTGGTTGTCAGAC5000                        AACACATGAGAAAGAAAAAGATGATGATGGTGGCAGTCTGAAGACGACAA5050                        TGAAAGATAGTCACGTAAGGCAGATTTTCTTCGATGGAGGTGAGACATCC5100                        GCTAATGAAACAGGATTAAGTAGCGGGGATGCAGACACTATTATGACTCC5150                        AACGACAAAGGAGGATAATCATGTTATAGACGTAATGGAGGAAAGGCGAA5200                        CTGATGAAGAGGCCAAAGGGCAAAGCGTTCATGAA5235                                       (2) INFORMATION FOR SEQ ID NO:36:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 5235 nucleotides                                                  (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                      TTCATGAACGCTTTGCCCTTTGGCCTCTTCATCAGTTCGCCTTTCCTCCA50                          TTACGTCTATAACATGATTATCCTCCTTTGTCGTTGGAGTCATAATAGTG100                         TCTGCATCCCCGCTACTTAATCCTGTTTCATTAGCGGATGTCTCACCTCC150                         ATCGAAGAAAATCTGCCTTACGTGACTATCTTTCATTGTCGTCTTCAGAC200                         TGCCACCATCATCATCTTTTTCTTTCTCATGTGTTGTCTGACAACCAAAC250                         GTTCCTTCTTTTGATGTTTCTTGTTGCTCAAAATTCATTTCCACTTCTCG300                         AACCATTCGATCCCATTTTTCTTGCTCCTCTTCGTTGAGAGGCACTATTT350                         TGTCAACAATTTCTTCTTCGTTTACAGCAACATCATCCTCATACGTAGTT400                         GTTATTGTACGCTCTTCCACAATAGGACCATTTTCCGAATCATGCACATG450                         TCGTTCAGTGCGCACAACTGTTCGAATTTCTGAAACTGCAACTTCCTCTT500                         CCTCTTCGTGGAATGACTGTTCTTTGGGTTCTTCTGGATCTCCAGGTGTC550                         CCATTATACCCTACTTGCTGAAAAAACGGTTCTTGTGCCAATCGCTGTTG600                         CGCCATTGTTACCTCGGCGTGACGACGTCTGTCGCCTGCTACCTCTAGCA650                         AAGTAGATGACAGAGTTATTGAGGGTCCAGAAATATGCGATACAGGTGTT700                         CCTTCTAAACCATCTAGCTTTTCAGCTCGATCCATTTCTCGTACAACATC750                         ATCACGTCCTATTCGCTGCAATGCTGATCGCAAAGCAACGGGCGTAGCTT800                         GTTCTTTCTTCAAAAATATCCAAATACGTAGAATAGTGACTGCTTCAAGA850                         CCAACTAGTTGATGTCGAACTTGTCGTATATCAATATCAGGTACTTCAAG900                         CGCTCGAGCTAACCGATGCCAATCAGCGCCAATTAATAGTGCAACATGTG950                         CTAGTGGCAAGTTATCAGGTTCAGCAGTTTCATGGAAAGCTCCAACGTAT1000                        TTCTCAGTCAAAGAAGCTTCCGAATAGAAGAGTTTTTTGGAAACCATCGG1050                        CTCCGGCCCAGTGTATTCCGGAAGAGTGATTGCAAGAGTACACACTGGCG1100                        TCTGCGGAGGTAAATTTTCGGCTCTCAATTTTGGTTCTTTCATAAATACT1150                        ATCCGGCCATCAGCTGCAGTTTCGTTGTCCGTGTGAGTGCGTATCTTTAC1200                        CATGAAAGCAAGACGATTTTCTTGGAATGGTAGAAAATAAAGAGAAAGTT1250                        GGTCACCACTCTTGGTTATTGGAAGAAGATTTCCAGAAAATTCCAAAAAC1300                        TGATGTCGCCCACTTAAGACTTCTACATCTTTCGATTTCGCAATTTCAAT1350                        GAAACGTTCTTGTTTTTCCAGGGTTTTATCTTCCCGATCATCAGTCATAC1400                        AAAACAATCTCAACTGTCCTTCGGCAGGAAAAGTTCGTCGAGCAAAAATA1450                        AGAAATTTAGCCATATAAGGAACTGCAATTGCTTCATTGTAAACTTCTTG1500                        TGCCATTCGTGCCGCATCTCGCGGAGTTTGGCAATCCATCAACCAAAATC1550                        GAGCAGAAACCGTAGTTGTAAATGAAACGTCCTCACCAGTAAATGTTAAC1600                        TGGGTAGTTCCAGTAATATCTTCCCACTGTGCAGGAGAAGAACCTCCAGT1650                        TTTACTGCAGAGTAAACGCAGCGTCGGTGGTTCCTGTCCTGGTTGGCCAC1700                        TATATTGTGTTAACATTCCTTTATTTGAGCTTTGTGGCAATGGTATGCAC1750                        AGCGTTATGGGCTTATGGAATTTGCGACGACGCGGTTCAACAGTTACAAT1800                        TGGAGAAACAGCGACTCTATTCCCATGTAAACGAGTGACTATCTCTTGTG1850                        GAACTGGCTGGGCTTGCACAGATACTTTGATCGTCTTAGTCAAGGAACCA1900                        TCCGGAAATATGGCCTGCACATGAGGAACAACTGAAGAGAGTATTACACC1950                        ACCTTCTGGACCAACACAGTGCACTTCTTGCCGCACACGAGTAACGACCG2000                        CGAAATACATTGGGAAATCATTGGTCAGGATACGCGTAATCCGTGATGTA2050                        TGCAAATCATCAAGTTGCGACAACTCTTCTGCATCAAACGATTCATTGAG2100                        CACCTCTTGTACAGCATCTTCTGTTGCTTCAAGCTGATGCTCTTTCCAAT2150                        GCTGCCCATCATCAGAACGCAAAATGACAATCTCTCTCTCTCGTCCACGA2200                        AGTGATGCAAAATGTGGTACTTCCAATATAACAGGGCCTAAGAATTTTGC2250                        TCCATGTGGTGCCATTTCAAGTATACGTGANGCGAGCGCTTCACCTTCAC2300                        TTAATGGTGGTGGATGCGCTAACTTGTCCTTTCCAAGGTATCTGCATGTG2350                        ACCCGTGTAGGTTGCGGCGCTTTCCTCGGTGGTATAATGATTCTGACACC2400                        GGAATGCCTACAACCACGCATTGCTCCTCCACGTGCATCCACCATAAACG2450                        AAATTAAAAAACTAGGTTGCATGGTAGTTCTCATTATCGTAACATTGTCA2500                        GGAAGCGATGCATCTAAGGAAGGATCGGCCATCGCTGTAGTAATTGGTTC2550                        ATGTTGTGCTTTACGAATTAAGGCATCCAAATCGCCATCCGTACCTTCCA2600                        CTTCCGGACTTCGTGACAATGTCGGTTCTGTGGCATGAATCAAATGTACA2650                        CCAGTTGAATCGTGCAAACCTTTTGTGAGGCTTTCTGAGAAATCATGAGC2700                        ATGAGCATTGGCTGTGATCTGATTATCTTCACCTTCATCTTCGGAATCGG2750                        AAAACATGGTTTCATTCATTGCTTCGGGATTCTGAGGTTTATATCTTTCT2800                        TCAGTAACGGTTGTGGTCTCCGTTATCACAGTAGTCTCGGTAATTGTTTT2850                        AAGCGCTTCAACCACGGATACATACCCTAGACGTTCAGCAATCGATAATG2900                        GAGTTTGTCCTGTCGATGTATGAACATTTGGACTTGCACCATGTTCCAAC2950                        AAGTAACGTACAACACTGTTATGCCCTTGCTGAGCAGCTTGATGCAGAGG3000                        AGTATAGGAAGCACGAGTAATAACTGAAACTCGTGCGCCATGCTCAATCA3050                        AGAAACGGACCATGTTTATTTGTCCAAAATGGCAAGCAACATGTAACGGC3100                        GTATATCCTGCTTTCGTTTTGGGATCAATGGCTGCGTTTTCTTTCACTAG3150                        TTCTTCTGCTACGCTCACACGATCCTCCTGTGCACATAAATGCATTGGTG3200                        TCAAGCCATTCCTTGCCTGAGCTCCAACTTTTGCTCCATTTTCAATTAAT3250                        AACGCAGCCATTTCGCGATGGCCCTCCTGGGCGGCAAGATGAAGTGGTGT3300                        AAAGCCAGCTTTGCTTTCAGCATTCGCATTTGCCTTATAATGAAGGAGAG3350                        TGCTAGCAATATCCATCTGATTCTTCTTCGCGGCAATATGTAAAGGAGTG3400                        TACCCATTCTTGGCAGCGGCATGTGCAGAAGCACCATTTTCTAGAAGTAA3450                        CAATGCTACCTTGTCGTTATTGTAATGTGCCGCTACATGCAGAGGTGTTA3500                        CCTGATTCTTGCCTTCAATGTCAACCGGTGTTCCTCGTTCTAGCAATGAT3550                        TTCGCGACCGGCAAATTGCCATACTTAGCAGCTAAATGCAACGGCGTAAA3600                        ACCCTTTTTCGTGAGCAGTGTCTTGTCGGTTCCATGATCCATCAATATTG3650                        CTGCCACTTCCTCTTGCCCCTCCTTGGCAGCAATATGAAGAGGAGTATAA3700                        AGATCTCTTGTGGCAGCATTTGGTGATGCATTAGCCTGCAGCAACAAAAT3750                        GACGATGTCGGTATTACCAAGACGTGATGCAATGTGCAGTGGAGTTTGTA3800                        GTTCACGAGCAGCAGCATCCACCTGTGCTCCATTACGCACCAAAACACGA3850                        ACAATGTCCGTTTGGTTTGCTCGTGCAGCTAAATGAAGAGGCGTTTCACC3900                        GCGTACTGTAGCCACATCTGCATTAGCACCTTGTTGTAGTAAATAGATGA3950                        CAATGTTTATAGCACCCATAAAAGCAGCGACATGCAGCGGTGAGAGACCG4000                        GATTCAGTAGTTGCTTCGATTGCAGCGTGGTATTTCAGTAGCAGTTCGAC4050                        AATTTTAATGCGATTTTTTTTGCAAGCGATATGCAGCGGTGTGAAGCCAT4100                        TGAGAGCTCGAGCATTCGGGTCAGCATTACGATCCAGCAAAAGTTTAGCG4150                        ACACGGACATGTCCGCAATGAGCAGCCACATGAAGAGGAGTGAGATAGTC4200                        AACAGTAACATCATCCACCTGTGCTGCCATATGTAAGGGAGCCAAACCAT4250                        TTTTTGTCTTAGCACTGATTGGAGCTCCTTTTTCAAGCAACAAATCAACA4300                        ACTTGATCATGACCTGAACGAGAAGCACAGTGTAATGGTGTTAGTAAATC4350                        ACGTGTGCGACAGTCAATTACGGCCCCATGAGCCAACAATAACGAAACCA4400                        TGTTTGTACGACCCCATTTTGTTGCAACGTGTAACGGACTTATGTTATGT4450                        CTCGCTTGGTAATTCACATTGGCTCCCTTTTCGAGTAGCAGTTGTGCTAC4500                        GTTCTCATTTCCATAGTGAGCGGCGATATGAAGCGGAGTAAAGCCGCTTT4550                        TCGAAGTCACATCCGAGTTATGCTCATTTTGAAGTAATAGCGTAGCTGCT4600                        TTCGTATCATCTTTTTTAGCAGCAATATGCAGTGCTGGCAAGCGCACTTT4650                        CCCGCGCGTGTCATTTTCAAGCAAAACAGCGACCACACGATCGTGACCTT4700                        GTTGCAAGGCAACTGCCAGTGGCGTAAAACCGTCTTCTGTACTTAAAGCT4750                        TGATTGGCATTGTGGGCAAGAAGATAGCGTACAACAGATTCGTGATTTTC4800                        TTGTGCAGCCATGTAAAGTGGTGTAAAACCGTTTAGTGATTGTACGTTAA4850                        CATTAGCACCATTTTCAACAAGTACTGTGACGATTAGTTCTTGTCCTGCC4900                        AATGATGCTATATGTAACGCTGTGTTACCCTTTCTAGTGGCAGCATCAAC4950                        ATCTGCTTTTCTTTTCAGAAGTTCGCGGACCACTTCATGATGACCTTCTT5000                        TGGAGGCCAGATGCAATGCATTAAGGCCATTCGCATTGCATGTGTTGATA5050                        TCGGTGCCCGAACGAAGTAGTTCAAGTACACGATCCAAATTTCCAGCTCT5100                        TGCTGCTCGTAAAAAACTTGCACTGCTCTCACCTTTGTTGGAATGTTGAC5150                        TGTTATCATCAGGAATTTGTTGTTGATGTTGTGAATCTTTTGGTTCTGCG5200                        GGCCAGCCACTTCCCTCGACTATAGGATTACTCAT5235                                       (2) INFORMATION FOR SEQ ID NO:37:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 908 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (ix) FEATURE:                                                                 (A) NAME/KEY: CDS                                                             (B) LOCATION: 1..906                                                          (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                      GATAATGTTACTGTTGACTATCTCACTCCTCTTCATGTGGCC42                                  AspAsnValThrValAspTyrLeuThrProLeuHisValAla                                    1510                                                                          GCCCACTGCGGACATGTCCGTGTCGCTAAGCTTCTGCTGGAT84                                  AlaHisCysGlyHisValArgValAlaLysLeuLeuLeuAsp                                    152025                                                                        CGTAATGCCGATTCAAATGCTCGGGCTCTCAATGGCTTCACA126                                 ArgAsnAlaAspSerAsnAlaArgAlaLeuAsnGlyPheThr                                    303540                                                                        CCGTTGCACATAGCTTGCAAAAAAAATCGCATTAAGGTTGTC168                                 ProLeuHisIleAlaCysLysLysAsnArgIleLysValVal                                    455055                                                                        GAACTGTTGCTGAAATATCATGCTGCCATCGAGGCAACTACA210                                 GluLeuLeuLeuLysTyrHisAlaAlaIleGluAlaThrThr                                    606570                                                                        GAATCCGGTCTGTCGCCGCTTCACGTCGCTGCTTTCATGGGT252                                 GluSerGlyLeuSerProLeuHisValAlaAlaPheMetGly                                    7580                                                                          GCTATAAACATCGTTATCTACTTACTGCAGCAGGGTGCTAAT294                                 AlaIleAsnIleValIleTyrLeuLeuGlnGlnGlyAlaAsn                                    859095                                                                        GCGAATGTGGCTACTGTACGCGGTGAAACACCTCTTCATTTA336                                 AlaAsnValAlaThrValArgGlyGluThrProLeuHisLeu                                    100105110                                                                     GCTGCACGAGCGAACCAAACCGATATTGTCCGTGTTTTGGTA378                                 AlaAlaArgAlaAsnGlnThrAspIleValArgValLeuVal                                    115120125                                                                     CGTAATGGAGCCCAGGTGGATGCCGCGGCACGTGAGCTACAA420                                 ArgAsnGlyAlaGlnValAspAlaAlaAlaArgGluLeuGln                                    130135140                                                                     ACACCATTACATATTGCATCACGTCTTGGCAATACTGATATC462                                 ThrProLeuHisIleAlaSerArgLeuGlyAsnThrAspIle                                    145150                                                                        GTTATCTTGTTGCTGCAGGCAGACGCATCACCAAATGCTGCT504                                 ValIleLeuLeuLeuGlnAlaAspAlaSerProAsnAlaAla                                    155160165                                                                     ACACGGGATCTCTACACTCTTCTTCATATTGCTGCCAAAGAG546                                 ThrArgAspLeuTyrThrLeuLeuHisIleAlaAlaLysGlu                                    170175180                                                                     GGACAAGAGGAGGTGGCAGCAATATTGATAGATCATGGTTCC588                                 GlyGlnGluGluValAlaAlaIleLeuIleAspHisGlySer                                    185190195                                                                     GATAAGACATTGCTTACCAAGAAAGGTTTTACACCGTTGCAT630                                 AspLysThrLeuLeuThrLysLysGlyPheThrProLeuHis                                    200205210                                                                     TTAGCTGCTAAATACGGCAATTTACCGGTAGCGAAATTATTG672                                 LeuAlaAlaLysTyrGlyAsnLeuProValAlaLysLeuLeu                                    215220                                                                        CTGGAACGAGGAACTTTGGTTGACATTGAAGGCAAGAACCAG714                                 LeuGluArgGlyThrLeuValAspIleGluGlyLysAsnGln                                    225230235                                                                     GTGACACCTTTGCATGTAGCAGCACATTATAATAACGACAAG756                                 ValThrProLeuHisValAlaAlaHisTyrAsnAsnAspLys                                    240245250                                                                     GTAGCGCTGCTGCTTCTAGAAAGTGGTGCTTCCGCACATGCC798                                 ValAlaLeuLeuLeuLeuGluSerGlyAlaSerAlaHisAla                                    255260265                                                                     GTTGCCAAGAATGGATATACTCCTTTGCATATTGCTGCAAAG840                                 ValAlaLysAsnGlyTyrThrProLeuHisIleAlaAlaLys                                    270275280                                                                     AAAAATCAGATGGATATTGCTAGCACTCTTCTTCATTATAGG882                                 LysAsnGlnMetAspIleAlaSerThrLeuLeuHisTyrArg                                    285290                                                                        GCAAATGCGAATGCTGAAAGCAAAGC908                                                 AlaAsnAlaAsnAlaGluSerLys                                                      295300                                                                        (2) INFORMATION FOR SEQ ID NO:38:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 302 amino acids                                                   (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                      AspAsnValThrValAspTyrLeuThrProLeuHisValAla                                    1510                                                                          AlaHisCysGlyHisValArgValAlaLysLeuLeuLeuAsp                                    152025                                                                        ArgAsnAlaAspSerAsnAlaArgAlaLeuAsnGlyPheThr                                    303540                                                                        ProLeuHisIleAlaCysLysLysAsnArgIleLysValVal                                    455055                                                                        GluLeuLeuLeuLysTyrHisAlaAlaIleGluAlaThrThr                                    606570                                                                        GluSerGlyLeuSerProLeuHisValAlaAlaPheMetGly                                    7580                                                                          AlaIleAsnIleValIleTyrLeuLeuGlnGlnGlyAlaAsn                                    859095                                                                        AlaAsnValAlaThrValArgGlyGluThrProLeuHisLeu                                    100105110                                                                     AlaAlaArgAlaAsnGlnThrAspIleValArgValLeuVal                                    115120125                                                                     ArgAsnGlyAlaGlnValAspAlaAlaAlaArgGluLeuGln                                    130135140                                                                     ThrProLeuHisIleAlaSerArgLeuGlyAsnThrAspIle                                    145150                                                                        ValIleLeuLeuLeuGlnAlaAspAlaSerProAsnAlaAla                                    155160165                                                                     ThrArgAspLeuTyrThrLeuLeuHisIleAlaAlaLysGlu                                    170175180                                                                     GlyGlnGluGluValAlaAlaIleLeuIleAspHisGlySer                                    185190195                                                                     AspLysThrLeuLeuThrLysLysGlyPheThrProLeuHis                                    200205210                                                                     LeuAlaAlaLysTyrGlyAsnLeuProValAlaLysLeuLeu                                    215220                                                                        LeuGluArgGlyThrLeuValAspIleGluGlyLysAsnGln                                    225230235                                                                     ValThrProLeuHisValAlaAlaHisTyrAsnAsnAspLys                                    240245250                                                                     ValAlaLeuLeuLeuLeuGluSerGlyAlaSerAlaHisAla                                    255260265                                                                     ValAlaLysAsnGlyTyrThrProLeuHisIleAlaAlaLys                                    270275280                                                                     LysAsnGlnMetAspIleAlaSerThrLeuLeuHisTyrArg                                    285290                                                                        AlaAsnAlaAsnAlaGluSerLys                                                      295300                                                                        (2) INFORMATION FOR SEQ ID NO:39:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 908 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                      GCTTTGCTTTCAGCATTCGCATTTGCCCTATAATGAAGAAGAGTGCTAGC50                          AATATCCATCTGATTTTTCTTTGCAGCAATATGCAAAGGAGTATATCCAT100                         TCTTGGCAACGGCATGTGCGGAAGCACCACTTTCTAGAAGCAGCAGCGCT150                         ACCTTGTCGTTATTATAATGTGCTGCTACATGCAAAGGTGTCACCTGGTT200                         CTTGCCTTCAATGTCAACCAAAGTTCCTCGTTCCAGCAATAATTTCGCTA250                         CCGGTAAATTGCCGTATTTAGCAGCTAAATGCAACGGTGTAAAACCTTTC300                         TTGGTAAGCAATGTCTTATCGGAACCATGATCTATCAATATTGCTGCCAC350                         CTCCTCTTGTCCCTCTTTGGCAGCAATATGAAGAAGAGTGTAGAGATCCC400                         GTGTAGCAGCATTTGGTGATGCGTCTGCCTGCAGCAACAAGATAACGATA450                         TCAGTATTGCCAAGACGTGATGCAATATGTAATGGTGTTTGTAGCTCACG500                         TGCCGCGGCATCCACCTGGGCTCCATTACGTACCAAAACACGGACAATAT550                         CGGTTTGGTTCGCTCGTGCAGCTAAATGAAGAGGTGTTTCACCGCGTACA600                         GTAGCCACATTCGCATTAGCACCCTGCTGCAGTAAGTAGATAACGATGTT650                         TATAGCACCCATGAAAGCAGCGACGTGAAGCGGCGACAGACCGGATTCTG700                         TAGTTGCCTCGATGGCAGCATGATATTTCAGCAACAGTTCGACAACCTTA750                         ATGCGATTTTTTTTGCAAGCTATGTGCAACGGTGTGAAGCCATTGAGAGC800                         CCGAGCATTTGAATCGGCATTACGATCCAGCAGAAGCTTAGCGACACGGA850                         CATGTCCGCAGTGGGCGGCCACATGAAGAGGAGTGAGATAGTCAACAGTA900                         ACATTATC908                                                                   (2) INFORMATION FOR SEQ ID NO:40:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 906 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                      GATAATGTTACTGTTGACTATCTCACTCCTCTTCATGTGGCCGCCCACTG50                          CGGACATGTCCGTGTCGCTAAGCTTCTGCTGGATCGTAATGCCGATTCAA100                         ATGCTCGGGCTCTCAATGGCTTCACACCGTTGCACATAGCTTGCAAAAAA150                         AATCGCATTAAGGTTGTCGAACTGTTGCTGAAATATCATGCTGCCATCGA200                         GGCAACTACAGAATCCGGTCTGTCGCCGCTTCACGTCGCTGCTTTCATGG250                         GTGCTATAAACATCGTTATCTACTTACTGCAGCAGGGTGCTAATGCGAAT300                         GTGGCTACTGTACGCGGTGAAACACCTCTTCATTTAGCTGCACGAGCGAA350                         CCAAACCGATATTGTCCGTGTTTTGGTACGTAATGGAGCCCAGGTGGATG400                         CCGCGGCACGTGAGCTACAAACACCATTACATATTGCATCACGTCTTGGC450                         AATACTGATATCGTTATCTTGTTGCTGCAGGCAGACGCATCACCAAATGC500                         TGCTACACGGGATCTCTACACTCTTCTTCATATTGCTGCCAAAGAGGGAC550                         AAGAGGAGGTGGCAGCAATATTGATAGATCATGGTTCCGATAAGACATTG600                         CTTACCAAGAAAGGTTTTACACCGTTGCATTTAGCTGCTAAATACGGCAA650                         TTTACCGGTAGCGAAATTATTGCTGGAACGAGGAACTTTGGTTGACATTG700                         AAGGCAAGAACCAGGTGACACCTTTGCATGTAGCAGCACATTATAATAAC750                         GACAAGGTAGCGCTGCTGCTTCTAGAAAGTGGTGCTTCCGCACATGCCGT800                         TGCCAAGAATGGATATACTCCTTTGCATATTGCTGCAAAGAAAAATCAGA850                         TGGATATTGCTAGCACTCTTCTTCATTATAGGGCAAATGCGAATGCTGAA900                         AGCAAA906                                                                     (2) INFORMATION FOR SEQ ID NO:41:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 906 nucleotides                                                   (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: cDNA                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                      TTTGCTTTCAGCATTCGCATTTGCCCTATAATGAAGAAGAGTGCTAGCAA50                          TATCCATCTGATTTTTCTTTGCAGCAATATGCAAAGGAGTATATCCATTC100                         TTGGCAACGGCATGTGCGGAAGCACCACTTTCTAGAAGCAGCAGCGCTAC150                         CTTGTCGTTATTATAATGTGCTGCTACATGCAAAGGTGTCACCTGGTTCT200                         TGCCTTCAATGTCAACCAAAGTTCCTCGTTCCAGCAATAATTTCGCTACC250                         GGTAAATTGCCGTATTTAGCAGCTAAATGCAACGGTGTAAAACCTTTCTT300                         GGTAAGCAATGTCTTATCGGAACCATGATCTATCAATATTGCTGCCACCT350                         CCTCTTGTCCCTCTTTGGCAGCAATATGAAGAAGAGTGTAGAGATCCCGT400                         GTAGCAGCATTTGGTGATGCGTCTGCCTGCAGCAACAAGATAACGATATC450                         AGTATTGCCAAGACGTGATGCAATATGTAATGGTGTTTGTAGCTCACGTG500                         CCGCGGCATCCACCTGGGCTCCATTACGTACCAAAACACGGACAATATCG550                         GTTTGGTTCGCTCGTGCAGCTAAATGAAGAGGTGTTTCACCGCGTACAGT600                         AGCCACATTCGCATTAGCACCCTGCTGCAGTAAGTAGATAACGATGTTTA650                         TAGCACCCATGAAAGCAGCGACGTGAAGCGGCGACAGACCGGATTCTGTA700                         GTTGCCTCGATGGCAGCATGATATTTCAGCAACAGTTCGACAACCTTAAT750                         GCGATTTTTTTTGCAAGCTATGTGCAACGGTGTGAAGCCATTGAGAGCCC800                         GAGCATTTGAATCGGCATTACGATCCAGCAGAAGCTTAGCGACACGGACA850                         TGTCCGCAGTGGGCGGCCACATGAAGAGGAGTGAGATAGTCAACAGTAAC900                         ATTATC906                                                                     (2) INFORMATION FOR SEQ ID NO:42:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                      CATCAATTTTTGGAATTTTCTGG23                                                     (2) INFORMATION FOR SEQ ID NO:43:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                      CGTTTACAGCAACATCATCCTC22                                                      (2) INFORMATION FOR SEQ ID NO:44:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                      GCACAACCAGTTCCGCAAGAAA22                                                      (2) INFORMATION FOR SEQ ID NO:45:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                      GGTTATTGGAAGAAGATTTCC21                                                       (2) INFORMATION FOR SEQ ID NO:46:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (ix) FEATURE:                                                                 (A) NAME/KEY: N = any nucleotide                                              (B) LOCATION: 9, 13                                                           (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                      CAYCARGCNGCNCARCARGGNCA23                                                     (2) INFORMATION FOR SEQ ID NO:47:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 23 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (ix) FEATURE:                                                                 (A) NAME/KEY: N = any nucleotide                                              (B) LOCATION: 3, 13, 16, 19                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                      GTNGAYGAYGTNACNGTNGAYTA23                                                     (2) INFORMATION FOR SEQ ID NO:48:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                      GGAATTTGCGACGACGCGGTTC22                                                      (2) INFORMATION FOR SEQ ID NO:49:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                      CAGGAAACAGCTATGAC17                                                           (2) INFORMATION FOR SEQ ID NO:50:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 24 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                      TGGAGTTTGTCCTGTCGATGTATG24                                                    (2) INFORMATION FOR SEQ ID NO:51:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                      GCTTTGCTTTCAGCATTCGCATTTGCC27                                                 (2) INFORMATION FOR SEQ ID NO:52:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 22 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                      GGTTTAATTACCCAAGTTTGAG22                                                      (2) INFORMATION FOR SEQ ID NO:53:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 27 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                      GTGAGATAGTCAACAGTAACATCATCC27                                                 (2) INFORMATION FOR SEQ ID NO:54:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 31 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                      CGCGGATCCGGCACAACCAGTTCCGCAAGAA31                                             (2) INFORMATION FOR SEQ ID NO:55:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 30 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                      CCGGAATTCTTATTCATGAACGCTTTGCCC30                                              (2) INFORMATION FOR SEQ ID NO:56:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                      CGCGGATCCGATGAGTAATCCTATAGTCGAGGG33                                           (2) INFORMATION FOR SEQ ID NO:57:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 35 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                      CCGGAATTCCGGTTACCCTAGACGTTCAGCAATCG35                                         (2) INFORMATION FOR SEQ ID NO:58:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 34 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                      CGCGGATCCGCGCGCACGTGGAGGAGCAATGCGT34                                          (2) INFORMATION FOR SEQ ID NO:59:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 36 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                      CCGGAATTCCGGTTATTCGTTGTCCGTGTGAGTGCG36                                        (2) INFORMATION FOR SEQ ID NO:60:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 37 nucleotides                                                    (B) TYPE: nucleic acid                                                        (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: primer                                                    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                      CGCGGATCCGCGCCAACTAGTTGGTCTTGAAGCAGTC37                                       (2) INFORMATION FOR SEQ ID NO:61:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                      GluSerSerAlaSerPheLeuArgAlaAlaArgAlaGlyAsn                                    1510                                                                          LeuAspArgValLeuGluLeuLeuArgSerGlyThrAspIle                                    152025                                                                        AsnThrCysAsnAla                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:62:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                      AsnGlyLeuAsnAlaLeuHisLeuAlaSerLysGluGlyHis                                    1510                                                                          HisGluValValArgGluLeuLeuLysArgLysAlaAspVal                                    152025                                                                        AspAlaAlaThrArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:63:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                      LysGlyAsnThrAlaLeuHisIleAlaSerLeuAlaGlyGln                                    1510                                                                          GluLeuIleValThrValLeuValGluAsnGlyAlaAsnVal                                    152025                                                                        AsnValGlnSerLeu                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:64:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                      AsnGlyPheThrProLeuTyrMetAlaAlaGlnGluAsnHis                                    1510                                                                          GluSerValValArgTyrLeuLeuAlaHisAsnAlaAsnGln                                    152025                                                                        AlaLeuSerThrGlu                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:65:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 29 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                      AspGlyPheThrProLeuAlaValAlaLeuGlnGlnGlyHis                                    1520                                                                          AspArgValValAlaValLeuLeuGluAsnAspThrArgGly                                    152025                                                                        Lys                                                                           (2) INFORMATION FOR SEQ ID NO:66:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                      ValArgLeuProAlaLeuHisIleAlaAlaLysLysAspAsp                                    1510                                                                          ThrLysAlaAlaThrLeuLeuLeuGlnAsnGluHisAsnSer                                    152025                                                                        AspValThrSerLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:67:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                      SerGlyPheThrProLeuHisIleAlaAlaHisTyrGlyAsn                                    1510                                                                          GluAsnValAlaGlnLeuLeuLeuGluLysGlyAlaAsnVal                                    152025                                                                        AsnTyrGlnAlaArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:68:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                      HisAsnIleSerProLeuHisValAlaThrLysTrpGlyArg                                    1510                                                                          ThrAsnMetValSerLeuLeuLeuAlaHisGlyAlaValIle                                    152025                                                                        AspCysArgThrArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:69:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                      AspLeuLeuThrProLeuHisCysAlaSerArgSerGlyHis                                    1510                                                                          AspGlnValValAspLeuLeuLeuGluLysGlyAlaProIle                                    152025                                                                        SerAlaLysThrLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:70:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                      AsnGlyLeuAlaProLeuHisMetAlaAlaGlnValAspAsp                                    1510                                                                          ValThrVal                                                                     15                                                                            (2) INFORMATION FOR SEQ ID NO:71:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                      AspTyrLeuThrProLeuHisValAlaAlaHisCysGlyHis                                    1510                                                                          ValArgValAlaLysLeuLeuLeuAspArgAsnAlaAspPro                                    152025                                                                        AsnAlaArgAlaLeu                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:72:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                      AsnGlyPheThrProLeuHisIleAlaCysLysLysAsnArg                                    1510                                                                          IleLysIleValGluLeuLeuLeuLysTyrHisAlaAlaIle                                    152025                                                                        GluAlaThrThrGlu                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:73:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                      SerGlyLeuSerProLeuHisValAlaAlaPheMetGlyAla                                    1510                                                                          IleAsnIleValIleTyrLeuLeuGlnGlnGlyAlaAsnAla                                    152025                                                                        AspValAlaThrVal                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:74:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                      ArgGlyGluThrProLeuHisLeuAlaAlaArgAlaAsnGln                                    1510                                                                          ThrAspIleValArgValLeuValArgAsnGlyAlaGlnVal                                    152025                                                                        AspAlaAlaAlaArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:75:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                      GluLeuGlnThrProLeuHisIleAlaSerArgLeuGlyAsn                                    1510                                                                          ThrAspIleValIleLeuLeuLeuGlnAlaAsnAlaSerPro                                    152025                                                                        AsnAlaAlaThrArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:76:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                      AspLeuTyrThrProLeuHisIleAlaAlaLysGluGlyGln                                    1510                                                                          GluGluValAlaAlaIleLeuMetAspHisGlyThrAspLys                                    152025                                                                        ThrLeuLeuThrLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:77:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                      LysGlyPheThrProLeuHisLeuAlaAlaLysTyrGlyAsn                                    1510                                                                          LeuProValAlaLysSerLeuLeuGluArgGlyThrProVal                                    152025                                                                        AspIleGluGlyLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:78:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                      AsnGlnValThrProLeuHisValAlaAlaHisTyrAsnAsn                                    1510                                                                          AspLysValAlaLeuLeuLeuLeuGluAsnGlyAlaSerAla                                    152025                                                                        HisAlaAlaAlaLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:79:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                      AsnGlyTyrThrProLeuHisIleAlaAlaLysLysAsnGln                                    1510                                                                          MetAspIleAlaSerThrLeuLeuHisTyrLysAlaAsnAla                                    152025                                                                        AsnAlaGluSerLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:80:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                      AlaGlyPheThrProLeuHisLeuAlaAlaGlnGluGlyHis                                    1510                                                                          ArgGluMetAlaAlaLeuLeuIleGluAsnGlyAlaLysVal                                    152025                                                                        GlyAlaGlnAlaArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:81:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                      AsnGlyLeuThrProMetHisLeuCysAlaGlnGluAspArg                                    1510                                                                          ValSerValAlaGluGluLeuValLysGluAsnAlaAlaIle                                    152025                                                                        AspProLysThrLys                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:82:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                      AlaGlyTyrThrProLeuHisValAlaCysHisPheGlyGln                                    1510                                                                          IleAsnMetValArgPheLeuIleGluHisGlyAlaArgVal                                    152025                                                                        SerValIleThrArg                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:83:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                      AlaSerTyrThrProLeuHisGlnAlaAlaGlnGlnGlyHis                                    1510                                                                          AsnSerValValArgTyrLeuLeuGluHisGlyAlaSerPro                                    152025                                                                        AsnValHisThrSer                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:84:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                      ThrGlyGlnThrProLeuSerIleAlaGluArgLeuGlyTyr                                    1510                                                                          ValSerValValGluAlaLeuLysThrIleThrGluThrThr                                    152025                                                                        ValIleThrGluThr                                                               30                                                                            (2) INFORMATION FOR SEQ ID NO:85:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 33 amino acids                                                    (B) TYPE: amino acid                                                          (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: protein                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Xaa = any amino acid                                            (B) LOCATION: 1, 3, 8, 11, 12, 15, 16,                                        17, 19, 20, 23, 24, 27, 28,                                                   30, 31, 32, 33                                                                (ix) FEATURE:                                                                 (A) NAME/KEY: Xaa = Val or Ala                                                (B) LOCATION: 18                                                              (ix) FEATURE:                                                                 (A) NAME/KEY: Xaa = Asn or Asp                                                (B) LOCATION: 29                                                              (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                      XaaGlyXaaThrProLeuHisXaaAlaAlaXaaXaaGlyHis                                    1510                                                                          XaaXaaXaaXaaXaaXaaLeuLeuXaaXaaGlyAlaXaaXaa                                    152025                                                                        XaaXaaXaaXaaXaa                                                               30                                                                            __________________________________________________________________________

What is claimed is:
 1. An isolated ankyrin nucleic acid moleculecomprises, wherein said nucleic acid molecule is a Dirofilaria or Brugianucleic acid molecule comprising a nucleic acid sequence selected fromthe group consisting of: a nucleic acid sequence selected from the groupconsisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ IDNO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ IDNO:13, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ IDNO:19, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ IDNO:26, SEQ ID NO:27, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ IDNO:32, SEQ ID NO:34, SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ IDNO:39, SEQ ID NO:40 and SEQ ID NO:41; and an allelic variant of anucleic acid sequence selected from the group consisting of SEQ ID NO:1,SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:8, SEQ IDNO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:14, SEQ IDNO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:21, SEQ IDNO:22, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ IDNO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:34, SEQ IDNO:35, SEQ ID NO:36, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:40 and SEQ IDNO:41.
 2. The ankyrin nucleic acid molecule of claim 1, wherein saidnucleic acid molecule is selected from the group consisting of aDirofilaria immitis ankyrin nucleic acid molecule and a Brugia malayiankyrin nucleic acid molecule.
 3. The ankyrin nucleic acid molecule ofclaim 1, wherein said nucleic acid molecule is selected from the groupconsisting of: a nucleic acid molecule selected from the groupconsisting of nDiAnk₉₃₇, nDiAnk₉₃₆, nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀,nDiAnk₁₂₂₈, nDiAnk₁₂₂₇, nDiAnk₅₇₃, nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆,nDiAnk₁₀₄₄, nDiAnk₅₅₀₃, nDiAnk₅₂₃₅, nBmAnk₉₀₈, and nBmAnk₉₀₆, and anallelic variant of a nucleic acid molecule selected from the groupconsisting of nDiAnk₉₃₇, nDiAnk₉₃₆, nDiAnk₁₀₂₉, nDiAnk₈₁₀, nDiAnk₆₀₀,nDiAnk₁₂₂₈, nDiAnk₁₂₂₇, nDiAnk₅₇₃, nDiAnk₉₁₁, nDiAnk₉₀₉, nDiAnk₁₀₉₆,nDiAnk₁₀₄₄, nDiAnk₅₅₀₃, nDiAnk₅₂₃₅, nBmAnk₉₀₈, and nBmAnk₉₀₆.
 4. Arecombinant molecule comprising an ankyrin nucleic acid molecule as setforth in claim 1 operatively linked to a transcription control sequence.5. A recombinant virus comprising an ankyrin nucleic acid molecule asset forth in claim
 1. 6. A recombinant cell comprising an ankyrinnucleic acid molecule as set forth in claim
 1. 7. A method of producingan ankyrin protein derived from Dirofilaria or Brugia, said methodcomprising culturing the cell of claim 6 in a culture media andisolating the ankyrin protein therefrom.
 8. An isolated ankyrin nucleicacid molecule selected from the group consisting of: a nucleic acidmolecule encoding a protein comprising an amino acid sequence selectedfrom the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:28, SEQ ID NO:33, andSEQ ID NO:38; and an allelic variant of a nucleic acid molecule encodinga protein comprising an amino acid sequence selected from the groupconsisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQ ID NO:15, SEQID NO:20, SEQ ID NO:23, SEQ ID NO:28, SEQ ID NO:33, and SEQ ID NO:38. 9.A recombinant molecule comprising an ankyrin nucleic acid molecule asset forth in claim 8 operatively linked to a transcription controlsequence.
 10. A recombinant virus comprising an ankyrin nucleic acidmolecule as set forth in claim
 8. 11. A recombinant cell comprising anankyrin nucleic acid molecule as set forth in claim
 8. 12. A method ofproducing an ankyrin protein derived from Dirofilaria or Brugia, saidmethod comprising culturing the cell of claim 11 in a culture media andisolating the ankyrin protein therefrom.
 13. An isolated ankyrin nucleicacid molecule encoding a fragment of a protein, said protein selectedfrom the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:12, SEQID NO:15, SEQ ID NO:20, SEQ ID NO:23, SEQ ID NO:28, SEQ ID NO:33, andSEQ ID NO:38, and allelic variants thereof, said fragment having theability to bind to an antibody against the protein from which it wasobtained.
 14. A recombinant molecule comprising an ankyrin nucleic acidmolecule as set forth in claim 13 operatively linked to a transcriptioncontrol sequence.
 15. A recombinant virus comprising an ankyrin nucleicacid molecule as set forth in claim
 13. 16. A recombinant cellcomprising an ankyrin nucleic acid molecule as set forth in claim 13.17. A method of producing an ankyrin protein derived from Dirofilaria orBrugia, said method comprising culturing the cell of claim 16 in aculture media and isolating the ankyrin protein therefrom.
 18. Anisolated ankyrin nucleic acid which encodes a protein that is at least75% identical to PDiAnk₁₇₄₅ (SEQ ID NO:33) and has the ability to bindto an antibody against PDiAnk₁₇₄₅.
 19. An isolated ankyrin nucleic acidmolecule which encodes a protein that is at least 90% identical toPBmAnk₃₀₂ (SEQ ID NO:38) and has the ability to bind to an antibodyagainst PBmAnk₃₀₂.